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NP1O5C<< c<6X9`("Courier NewTTd6X@DQ@<6X9`("Courier NewTTXXx6X@DQX@<6X9`("Courier NewTTZ6X@DQ@<6X9`("Courier NewTT6X@DQ@<6X9`("Courier NewTT- -   $    5    ݀X&XXX&,&XX&Muchofthematerialinthischapterisderivedfromacommissionedpaperpreparedforthe  NationalBioethicsAdvisoryCommissionbyCourtneyS.Campbell,DepartmentofPhilosophy,OregonStateUniversity,titled ReligiousPerspectivesonHumanCloning.\  `$Times NewRoman4#)2Quick 1.J  .0 J($    UC:\WINDOWS\TEMPLATE\STANDARD.WPT  ^ - -- -\  `*Times New RomanTT k  $    1    &$&XXInsupportofitsanalysis,NBACcommissionedapaperwrittenbyDanBrock,Brown  University,titled CloningHumanBeings:AnAssessmentoftheEthicalIssuesProandCon.Someofthematerialinthischapterisderivedfromthatpaper.   $    2    &$&XXThereisoneargumentthathasbeenusedbyseveralcommentatorstounderminetheapparent  significanceofpotentialharmstoachildcreatedthroughsomaticcellnucleartransfer(Chadwick1982;Robertson1994,1997;Macklin1994).Thepointderivesfromageneralproblem,calledthenon-identityproblem,posedbythephilosopherDerekParfitandnotoriginallydirectedtohumancloning(Parfit1984).Thisviewarguesthatalltheproblemsofhavingbeenbornviasuchcloningarenotnetharmstotheresultingchildbecausetheyarenotworsethannolifeatall.Parfitdoesnotaccepttheaboveargumentassound.Instead,hebelievesthatifonecouldhaveadifferentchildwithouttheseburdens(forexample,byusingadifferentmethodofreproduction)thereisasstrongamoralreasontodoso(Brock1995). h  $    3    ݀&$&XXMoralphilosophersthinkaboutpersonhoodwhentheyconstructanddeploytheirviewsof  humanchoiceandmoralagency.ForKantians,personhoodisaboutfreewillandreason.FromthepointofviewofKantianmoralpersonality,allofusareidenticalaspersons.Philosophersofmindthinkaboutpersonhoodwhentheytrytofigureoutwhatconstitutespersonalidentity.Formanyofthesephilosophers,personalidentitymeanshavingacontinuouslifestorythatincorporatesapastandafutureforoneself.Fromthepointofviewofpersonalidentity,allofusaredifferent,unique,aspersons.Psychoanalyststhinkaboutpersonhoodwhentheyrelatetheconstantsofhumanlifeanddevelopmenttobroadpersonalitystructures.Fromthepsychoanalyticpointofview,eachofusmanifeststhesamedynamicpersonalitystructures,yetnotwoofusdosoinexactlythesameway;weareallthesameandalsoalldifferent.Welfarerightsactivistsandhumanrightsactivistsmaythinkaboutpersonhood:whatistheminimumofnecessaryresourcesforafullyhumanlife?Somemedicalethiciststhinkaboutpersonhoodwhiletryingtodecideatwhatpointdoeslifeceasetobeahumanlifeworthliving?Politicaltheoristsattimesthinkaboutpersonhoodinthecontextoftryingtounderstandwhatarethebasicsofindividualitythatthestateshouldrecognizeorunderwrite?Parentsthinkaboutpersonhood:whatpartdoIplayinmakingpossiblethefullestkindofhuman-nessformychildren?(Radin,1995).   $    4    &$&XXԀ Kantianethicalthought,writesRadin, distinguishesmorallybetweenpersonsandobjects.  Rationalbeingspossessingfreewill(persons)areautonomous;themorallawrequiresthatpersonsbetreatedasends,notmeans.Objectsinthenaturalworldthatarenotrationalbeingspossessingfreewillarenotpersons,andmayappropriatelybeusedasmeansbypersons.Kant'sviewrequiresthatpersons,moralagents,notbetreatedasobjects,manipulatedatthewillofpersons.KantpresentedhisbasicprinciplesofethicsinImmanuelKant,GroundworkofTheMetaphysicsofMorals(1785),translatedby . B H.J.PatoninTheMoralLaw(1948).[MargaretRadin, ReflectionsonObjectification,65Southern   CaliforniaLawReview341(November1991),atfootnote4]H       5    &$&XXBranzburgv.Hayes,408U.S.665,705(1972).Similarly,theSupremeCourtstatedinMeyerv.  NebraskathattherighttolibertyguaranteedbytheFourteenthAmendmentencompassedfreedomto  "acquireusefulknowledge...andgenerallytoenjoythoseprivilegeslongrecognizedatcommonlawasessentialtotheorderlypursuitofhappinessbyfreemen."      6    &$&XXHenleyv.Wise,303F.Supp.62(N.D.Ind.1969).C:\WINDOWS\TEMPLATE\STANDARD.WPT\  `*Times New RomanTTC\  P6QP\  `*Times New RomanTTXXP\  P6QXP .',text   s   Ua, b, c,Level 1Level 2Level 3Level 4Level 56#*4Quick a) o  )0 oH0%4heading 1heading 1&            '  U(3$ !    0  .( 9Z+8Courier 10 Pitch Regular<:Print From:?z*??%2A`Arial?/5  H/XXX?%2A`Arial?C:\WINDOWS\TEMPLATE\STANDARD.WPT- -K Kdyyyy'dxdC:\WINDOWS\TEMPLATE\STANDARD.WPT<:Print ReverMz~y6& dM &&&?%2A`Arial?/5  H/ XXX?%2A`Arial?- - d<:Reply/ForwarMz~y6& dM ?%2A`Arial?/5  H/ XXX?%2A`Arial?- -&$1Azy*A?%2A`Arial?/5  H/XXX?%2A`Arial?TABLE A$35;AGMU]e2I.A.1.a)(1)(a)(i)(a)0MyAArial BoldC:\WINDOWS\TEMPLATE\STANDARD.WPTX Xd# e37=CIQYag1.a.i.(1)(a)(i)1)a)'dxd%2A`Arialappendix-  A  $    1    ManyofthedefinitionswereexcerptedfromtheNationalInstitutesofHealthReportof  TheHumanEmbryoResearchPanel(Washington,DC:U.S.GovernmentPrintingOffice,1994).         H w@TABLE Aw@TABLE A   $    12    ݛX7KXXX&&XX7K#&$&&&E#&&&&$"[I]nordertobelegitimate,theState'sinterest...mustbesecular;consistentwiththeFirst  AmendmenttheStatemaynotpromoteatheologicalorsectarianinterest.PlannedParenthoodofSoutheasternPennsylvaniav.Casey,112S.Ct.2791,120L.Ed2d674,739(1992)(Stevens,J.concurringinpartanddissentinginpart).SeealsoThornburghv.AmericanCollegeofObstetriciansandGynecologists,476U.S.747,778(1986)(Stevens,J.concurring);seegenerallyWebsterv.ReproductiveHealthServices,492U.S.490,563-572(1989)(Stevens,J.,concurringinpartanddissentinginpart).  Whenappliedtoethicaldecisionmaking,onephilosophernotes:"Morality'sambitionis,oratleastoughttobe,toprovideasystemofconductunderwhicheveryonecanlivewithasenseofmutualϢjustifiability.Thisfollowsfromtheconditionsofpoliticallegitimacy.Wedonotliveinatheocracy,wheresomepeoplearethoughttohaveaprivilegedanddirectlinetomoraltruth."(Nagel1995)#&$&&&#&&&&$   $    8    ݛX7KXXX&&XX7KInmanystates,thewomanwhogivesbirthisconsideredtobethelegalmotherandherhusband  thelegalfatherofanyresultingchild.UnderstatutesinArizonaandUtah,thisholdstrueevenwhenthesurrogateisgestatinganembryowithnogeneticrelationshiptoher.OnlyinFlorida,NewHampshire,NorthDakotaandVirginiadocourtapprovedgestationalsurrogacyarrangementsresultintheintendedrearingparents"notthesurrogate"beingviewedasthelegalparents.#&$&&&D#u&{+&&&$#uX'X+&u&{c#   $    9    ݛX7KXXX&&XX7KThelatteroftenwillhaverights(eventhoughhehasnobiologicalconnectiontothechild)based  onthecommonlawpresumptionthatifawomangivesbirthwithinmarriage,herhusbandisthechild'slegalfather,orinsomestates,basedonspecificstatutesholdingthatthesurrogateandherhusbandarethelegalparentsofachildshehasgestatedregardlessoftheirgeneticcontribution.#X7KX&&c#Ԁ#XXXX7KD#uX'XXX#XX'XuXh#X7KXXX&&XX7KSee,e.g.,Ariz.Rev.Stat. n 25218(1996).#X7KX&&##XXXX7K#uX'XXX- - q  $    15    ݛX7KXXX&&XX7KLifchezv.Hartigan,735F.Supp.1361(N.D.Ill.),affdwithoutopinion,subnom.,Scholbergv.  ТLifchez,914F.2d260(7thCir.1990),cert.denied,111S.Ct.787(1991).  $    11    ݛX7KXXX&&XX7KSee,e.g.,Curlenderv.BioScienceLaboratories,165Cal.Rptr.477(Ct.App.1980).  $    2    X7KXXX&&XX7Kԛ42U.S.C.A.Sec.263a1etseq  $    3    X7KXXX&&XX7Kԛ45C.F.R.Part46 '  $    7    ݛX7KXXX&&XX7KIfcloningisconsideredtobeaformoffertilization,questionsariseregardingwhetherstatelaws  settingstandardsforwhomayperforminvitrofertilizationwillcoverthepractice.Certainlawsgoverning  reporting,thequalificationsofpersonnel,andsoforth,willbeapplicabletoresearchers.ANewHampshirelaw,forexample,requirescounselinginadvanceofinvitrofertilizationandlimitsthe n proceduretoparticipantsoverage21 C ׀(which,ifappliedtocloning,mightprohibittheuseofDNAfromaminorchild).Pennsylvaniahasareportingrequirementwhichmandatesthatanyoneperforminginvitro  . fertilizationfilequarterlyreportswiththeDepartmentofHealthdescribingsuchfactsasthenumberofembryosdestroyedanddiscardedandthenumberofwomeninwhomembryosareimplanted.(  C ׀Louisianaslawrequiresthatinvitrofertilizationshallonlybeundertakenbypractitionersandfacilitiesmeetingthe   standardsoftheAmericanCollegeofObstetriciansandGynecologists(ACOG)andtheAmericanFertilitySociety(AFS)(currently,theAmericanSocietyforReproductiveMedicine).La.Rev.Stat.Ann.9:128(West1991).#&$&&&D#u&{+&&&$#uX'X+&u&{c#   $    6    X7KXXXԛ&&XX7KTenstateshavelawsregulatingresearchand/orexperimentationonconceptuses,embryos,fetuses  orunbornchildrenthatusebroadenoughlanguagetoincludeearlystageconceptuses.Fla.Stat.Ann.390.001(6)(West1993);La.Rev.Stat.Ann.9:121etseq.(West1991);Me.Rev.Stat.Ann.tit.22,  1593(West1992);Mass.Gen.LawsAnn.ch.112,12J(West1996);Mich.Comp.LawsAnn.333.2685etseq.(WestSupp.1997);Minn.Stat.Ann.145.421(West1989);N.D.Cent.Code1402.2 DX 01(1991);N.H.Rev.Stat.Ann.168B:15(Supp.1996);Pa.Cons.Stat.3216(WestSupp.1996);R.I.Gen.Laws11541(1994).#X7KX&&c#   $    4    &$&XX&&&&$ StatementofthePresidentonNIHRecommendationsRegardingHumanEmbryoResearch,  U.S.Newswire(Dec.2,1994). V  $    14    ݛ[XXH[#X7KXHH#&&XX7KPlannedParenthoodv.Casey,505U.S.833,112S.Ct.2791,2810(1992).#X7KX&&#HXX7K   $    20    ݛX7KXXX&&XX7KEmmaThompson, GermansandFrenchPressforWorldwideBanonHumanCloning,The  Herald(Glasgow),April30,1997,p.14.   $    13    ݛX7KXXX&&XX7KSee,e.g.,Griswoldv.Connecticut,381U.S.379(1965);Eisenstadtv.Baird,405U.S.438  (1972).#&$&&&E#&&&&$ԀEarlydecisionsprotectedthemarriedcouplesrighttoprivacytomakeprocreativedecisions,but  laterdecisionsfocusedonindividualsrightsaswell.TheU.S.SupremeCourt,inEisenstadtv.Baird,  stated, [i]ftherightofprivacymeansanything,itistherightoftheindividual,marriedorsingle,tobe n freefromunwarrantedgovernmentalintrusionintomatterssofundamentallyaffectingapersonasthedecisionwhethertobearorbegetachild.Eisenstadtv.Baird,405U.S.438,453(1972).#&$&&&8#&&&&$   $    21    ݛX7KXXX&&XX7KԢGileTremlett, TwentyEuropeanCountriesSignInternationalConvention,TheTimes(U.K.),  April5,1997.  $    5    X7KXXX&&XX7KP.L.10491andP.L.104208.  $    22    ݛX7KXXX&&XX7K HealthAgencySaysCloningofHumansUnacceptable,ChicagoTribune,May15,1997.  $  X  Ӏ  43    .0 N.H.Rev.Stat.Ann.168B:13(Supp.1993)  $  X  Ӏ  44    ݛ.0 18Pa.Cons.Stat.Ann.3213(e)(1983).   $    16    ݛX7KXXX&&XX7KTheapplicabilityofMedicare(whichgenerallypaysforthecareofpersonsaged65orolder)  maynotbeapparent,butwiththeadventofpostmenopausalpregnancyviahormonalmaintenance,Medicareunexpectedlybecameapublicinsurerwithatleasttheoreticalobligationstopayforpregnancycare.Furthermore,evenifthefemalepartnerisnotcoveredbyMedicare,themalepartner,fromwhomthesomaticcellnucleusmightbeobtained,couldbeoldenoughtobeaMedicarebeneficiary.  $    18    X7KXXX&&XX7K GlobalGroupUrgesaVoluntaryBanonHumanCloning,ChicagoTribune,May12,1997,p.  16. n  $    1    ݛX7KXXX&&XX7KԀTosupporttheCommissionsreview,acommissionedpaper, TheCurrentandFutureLegal  StatusofCloning,waspreparedbyLoriAndrews,ChicagoKentCollegeofLaw.Inaddition,NBACcommissionedareviewofresearchmoratoria, DoResearchMoratoriaWork?preparedbyRobertM.CookDeegan,andareviewofinternationalresponses, Cloning:AnInternationalComparativePerspective,preparedbyBarthaKnoppers,UniversityofMontreal. "  $    10    &$&XX&&&&$See,e.g.,LosAngelesTimes,February25,1997,page6, Next,ReallyProlificCows:Scientists  CloneaSheep,butWeNeedn'tFrettheDoomsdayScenarios;TheNewYorkTimes,February25,1997,  SectionA;page26; CloningforGoodorEvil;TheHoustonChronicle,February25,1997,Outlook;   page19, Dolly'sbirthisfathertosomeworryingmusings,OtisPike;TheRecord,February25,1997,  pageL10, OfSheepandMen;BeforeBuildingaBetterBeast,ThinkTwice;TheSanDiego  Union-Tribune,February25,1997,pageB-6, Amazingbreakthrough:Cloningofsheephasremarkable  implications;WallStreetJournal,February25,1997,SectionA;page22, Review&Outlook:Listening n totheLamb;TheArizonaRepublic,February26,1997,pageB4, CloningQuestion;TheMysteriesof DX Life;TheFloridaTimes-Union,February26,1997,pageA10, Noneedforpanic;MiamiHerald,  . February26,1997,SectionA;page16, God'sWork;Man'sHands;TheMorningCall,February26,   1997,pageA16, 'Dolly'OpensNewVistasForMankind;St.PetersburgTimes,February26,1997,page   Т14A, Rulesforcloningneeded;TheBuffaloNews,February27,1997,page2B, ReadyorNot,Cloning   HasArrived;Don'tLoseTimeBanningitinHumans;DaytonDailyNews,February27,1997,page1a, r    AnimalCloningCallsforHumanRestraint;PhiladelphiaInquirer,February27,1997,page19, Don't H \  BeTooHastyWithLawsonCloning,byJamesKGlassman;TheSanFranciscoExaminer,February27, 2  1997,pageA20, HelloDolly:Thecloningofalambfromasheepcellopensupaneweraofnervousjokes,profoundquestionsandathleticopportunity;TheAugusta(Ga.)Chronicle,February28,1997,   pageA4, BanHumanCloning;TheStateJournal-Register(Springfield,IL),March2,1997,page16,    Cloningofsheepholdsremarkableimplications;TheBaltimoreSun,March3,1997,page8A, Moreof v  youandme?;Hello,Dolly:Replicatingasheepraisesconcernsaboutcloninghumans;TheIndianapolis L`  News,March4,1997,pageA6, Wolvesinsheep'scloning;TheSpokesman-Review(Spokane,WA), "6 March7,1997,pageB6, CloningTemptsOurDarkerSides;BanResearch;WeWon'tResisttheUrgetoTurnHumansintoInstruments,D.F.Oliveria;TheSpokesman-Review(Spokane,WA),March7,1997,  pageB6, CloningOffersHope,NotEvil;Don'tBeAfraid;CloningResearchOffersHopetoSolveGeneticMysteries,RebeccaNappi;TheTimes-Picayune,March10,1997,pageB6, CloningBegets z Questions;DaytonDailyNews,March10,1997,page6A, FearofClonesItselfaThreat;TheOrange Pd CountyRegister,March10,1997,pageB06, Vitalquestions;LosAngelesTimes,March13,1997,page &: 8, Don'tRushAnti-cloningLaws;ConcernsAreReal,butLegislationNeedsExpertInput;The  NashvilleBanner,March19,1997,pageA8, Frist'snoteofcaution;Don'tbetoohasty,hesays,topass  lawoncloning;TheNation,March24,1997,No.11,Vol.264;Pg.4;ISSN, Irreplaceableewe;cloning  ofasheep;Editorial,Hubbard,Ruth;TheNewYorkTimes,April1,1997,page22, Cloningasan ~ Anticlimax,PhilipM.Boffey;InformationBankAbstracts,WallStreetJournal,May2,1997,page14, Th  WillCloningBegetDisaster?#&$&&&c#&&&&$   $    17    X7KXXX&&XX7KToreceiveinputonscientificandprofessionalsocietyviewsaboutcloningofhumanbeings,  NBACcommissionedtheCriticalTechnologiesInstituteofRANDtorequestinformalinputfromrelevantorganizations,ofwhich32responded. ViewsofScientificSocietiesandProfessionalAssociationsonHumanNuclearTransferCloningResearch,byElisaEiseman,May1997.  $    19    X7KXXX&&XX7KSeetranscriptsofNBACHumanSubjectsSubcommitteemeeting,December16,1996. !  ݛXBXXXxrxXXB   `  #XBXxxr?#XXBCLONING      `  HUMAN   `  BEINGS#XBXΦ#xrxXXB P  #xxxxr##X XxxX#   `  ReportandRecommendationsofthe   `  NationalBioethicsAdvisoryCommission   `  Rockville,Maryland   `  June1997     XfXXX  X XfCONTENTS#r  ##X X r #   A- ) xdE0xA  0   (#(# 0  LetterofTransmittaltothePresident  (#(#    LetterfromthePresident     NBACRoster  StaffRoster  Acknowledgments 0 `   ` (#` (# ExecutiveSummary   h      p     i C  0 `  ` (#` (# 1.Introduction   h      p     1 #s  2.TheScienceandApplicationofCloning0  0p(#(#0p(#p(#13c (#(# 3.ReligiousPerspectives h      p   39 S 4.EthicalConsiderations h      p   63 C 5.LegalandPolicyConsiderations     p   87 3 6.RecommendationsoftheCommission   p   107 # Ѐ̜ r XX Appendices  #X X r# A.Glossary    h      p   1  B.ListofSpeakers   h      p   4  C.ListofCommissionedPapers     p   5  0   (#(# 0  0` (#(#  ` (#` (#   XXXX oXX#XXo7 ##X XXX #  k(#(   ?T̜June9,1997ThePresidentTheWhiteHouseWashington,D.C.20500̀DearMr.President:OnFebruary24,1997,inthewakeoftheannouncementthatscientistsinScotlandapparentlyhadsucceededincloninganadultsheep,youaskedtheNationalBioethicsAdvisoryCommissiontoreviewthelegalandethicalissuesassociatedwiththeuseofthistechnologyandtoreportbackwithinninetydayswithrecommendations.Aweeklateryouinstructedtheheadsofexecutivedepartmentsandagenciesthat"nofederalfundsshallbeallocatedforcloningofhumanbeings"therebyensuringthatprecipitousstepswouldnotbetakenwhiletheCommissionwasstudyingthesubject.Inthisshortinterval,wehavemadeeveryefforttoconsultwithethicists,theologians,scientists,physicians,andothercitizenswithinterestsandconcernsinthisarea.Moreover,wehaveinvitedinputsfortheCommission'sconsiderationfromasbroadacross-sectionofthecommunityastimeallowed.Further,recognizingthatscienceandmedicineareinternationalactivitieswithoutstandinginvestigatorsandfacilitiesinmanynations,wehaveattemptedtoreviewrelevantpoliciesandproposalswithrespecttohumancloninginothercountries.However,wedonotviewitasessentialtofollowothersinthisareaunlesswefindtheirproposalscompelling,sincewehavedifferentpoliticalandculturaltraditions.Inthisreport,weaddressaveryspecificaspectofcloningnamelywheregeneticmaterialwouldbetransferredfromthenucleusofasomaticcellofanexistinghumanbeingtoanenucleatedhumaneggwiththeintentionofcreatingachild.Wedonotrevisiteitherthequestionofthecloningofhumansbyembryosplittingortheissuessurroundingembryoresearch.Thelatterissuehas,ofcourse,recentlyreceivedcarefulattentionbyaNationalInstitutesofHealthpanel,theAdministration,andCongress.Notsurprisingly,wehavediscoveredthatthepotentialabilitytoclonehumanbeingsthroughthesomaticcellnucleartransfertechniquesraisesawholehostofcomplexanddifficultscientific,religious,legalandethicalissues--bothnewandold.Indeed,theCommissionitselfisunabletoagreeatthistimeonalltheethicalissuesthatsurroundtheissueofcloninghumanbeingsinthismanner.Itseemscleartoallofus,however,giventhecurrentstageofscienceinthisarea,thatanyattempttoclonehumanbeingsviasomaticcellnucleartransfertechniquesisuncertaininitsprospects,isunacceptablydangeroustothefetusand,therefore,morallyunacceptable.Atpresent,moralconsensusonthisissueshouldbeeasilyachieved.Furthermore,thecontinuing -@), ?TH  Page2ThePresidentcontroversyoverthesocialandethicalissuesraisedbythisnewdevelopmentrequiremoretimefordeliberationandtheaccumulationofnewscientificdata.Wethereforerecommendthatthecurrentmoratoriumonattemptstocreatechildreninthismannerbecontinuedandthatyouimmediatelyaskforvoluntarycomplianceintheprivatesectorwhilefederallegislationbanningtheuseofthesetechniquesforcreatingchildrenisformulatedandconsidered.Whilewehavebeenabletoagreeonthisandcertainotherrecommendedactions,wefeelquitestronglythatmostofthelegalandmoralissuesraisedcanonlyberesolved,eventemporarily,byagreatdealmorewidespreaddeliberationandeducation.Thistypeofnationaldiscussionisespeciallynecessaryinourdemocraticandpluralisticsocietyforthefollowingreasons:thereisnouniversallyacceptedethicaltheory;Americansholdvariousreligiousandmoralperspectivesontheseissues;conflictingvaluesareatstake;Americansdifferontheimportanceandmeaningofparticulartraditions;tolerance(agreeingtodisagree)governswideareasofournationallife;andgivenourhistoricaltraditions,wearestronglyinclinedtoleavetotheindividualconsciencethoseactsthatdonotharmothersandonwhichthereisnomoralconsensus.Asaresult,wemustcontinuetobuildourunderstandingofthewidespreadpublicconcernthathasbeengeneratedbytheserecentdevelopments.Someofthisconcerncanbeexplainedbyaninadequateunderstandingoftheissues--sometimesevenconfusingscienceandsciencefiction.Thismatter,however,canbeaddressedovertimethroughfurtherpubliceducation.Otherconcerns,however,runmuchdeeperandrangefromtheimplicationsforparticularfaithcommitments,toviewsregardingtheappropriatesphereforhumanaction,toconcernsregardingthefutureofthefamily,tocumulativeapprehensionsabouttherealnetbenefitofarapidlyadvancingtechnologythatsomebelieveistooaggressivelypushingasideimportantsocialandmoralvalues.Aswemoveaheadtothenextstageofournationaldiscussion,theseareamongthemanyissuesthatneedtobethoughtfullyaddressed.Finally,whileourspecificrecommendationsincludecontinuingthemoratoriumyouannouncedinFebruaryofthisyear,andacallforaspecificfederallegislation,thereportalsoincludesimportantsectionsoutliningthescientific,religious,ethicalandlegalissuesthatareraisedbythesenewscientificdevelopments.Itisourhopethatthesematerials,byclarifyingcertainissuesandhighlightingothers,willformausefulinitialbasisfortheongoingdeliberationsandeducationaldialoguesthatwebelievearesoessential.WehavebeenimpressedbythedifficultiescausedbythelackofknowledgeaboutgeneticsandthescienceinvolvedincloningrevealedinthepublicandmediaresponsestothecloningofDollythesheep.Webelieve,therefore,thatthefederalgovernmentshouldcontinuetoactivelyencouragepubliceducationinthisareaofsciencesothataspublicdeliberationtakesplaceitisasinformedaspossible.IwouldliketotakethisopportunitytothankalltheCommissionersandourverydedicatedstafffortheintensityanddepthoftheircommitmenttothetaskthatyouassignedtous.̀Sincerely,̛̜HaroldT.Shapiro .0*- Л` X XXX  j@| | THEWHITEHOUSE  @ZZWASHINGTONFebruary24,1997Dr.HaroldShapiroChairNationalBioethics̀AdvisoryCommissionSuite3C016100ExecutiveBoulevardBethesda,Maryland208927508DearDr.Shapiro:Asyouknow,itwasreportedtodaythatresearchershavedevelopedtechniquestoclonesheep.Thisrepresentsaremarkablescientificdiscovery,butonethatraisesimportantquestions.Whilethistechnologicaladvancecouldofferpotentialbenefitsinsuchareasasmedicalresearchandagriculture,italsoraisesseriousethicalquestions,particularlywithrespecttothepossibleuseofthistechnologytoclonehumanembryos.Therefore,IrequestthattheNationalBioethicsAdvisoryCommissionundertakeathoroughreviewofthelegalandethicalissuesassociatedwiththeuseofthistechnology,andreportbacktomewithinninetydayswithrecommendationsonpossiblefederalactionstopreventitsabuse.Sincerely,̜BillClinton#X XXX $# `'"%   P(#& hH`    XfXXX ENationalBioethicsAdvisoryCommission#X XX Xf[)#X XXX ԛ@@*   L) XX   HaroldT.Shapiro,Ph.D.Chair   PresidentPrincetonUniversity̜Princeton,NewJerseyQ*0   ^#^# М (X^#^#(PatriciaBacklar   SeniorScholarCenterforEthicsinHealthCareOregonHealthSciencesUniversityPortland,Oregon̜*,XFXX*SeniorResearchAssociateDepartmentofPhilosophyPortlandStateUniversityPortland,Oregoǹ̜ ArturoBrito,M.D.  Pp  AssistantProfessorofClinicalPediatricsUniversityofMiamiSchoolofMedicine̜Miami,Florida AlexanderMorganCapron,LL.B.  8X МHenryW.BruceProfessorofLawUniversityProfessorofLawandMedicineCoDirector,PacificCenterforHealthPolicyandπEthicsUniversityofSouthernCalifornia̜LosAngeles,Californià   EricJ.Cassell,M.D.  x МClinicalProfessorofPublicHealthCornellUniversityMedicalCollege̜NewYork,NewYork̜ R.AltaCharo,J.D.  `# AssociateProfessorofLawand̀MedicalEthicsSchoolsofLawandMedicineUniversityofWisconsin̜Madison,Wisconsin̜ JamesF.Childress,Ph.D.  " * KyleProfessorofReligiousStudiesProfessorofMedicalEducationDepartmentofReligiousStudiesUniversityofVirginia̜Charlottesville,Virginia DavidR.Cox,M.D.,Ph.D.  P(p&1 МProfessorofGeneticsandPediatricsDepartmentofGeneticsStanfordUniversitySchoolofMedicine̜Stanford,California RhetaughGravesDumas,Ph.D.,R.N.  - +7 МViceProvostforHealthAffairsLucilleColeProfessorofNursingTheUniversityofMichigan̜AnnArbor,Michigan̜  0/< ЇEzekielJ.Emanuel,M.D.,Ph.D.  < AssociateProfessorofMedicalEthicsDepartmentofSocialMedicine̜HarvardMedicalSchoolBoston,Massachusetts LaurieM.Flynn  0 P B ExecutiveDirectorNationalAllianceforthèMentallyIllArlington,Virginia CarolW.Greider,Ph.D.  H SeniorStaffScientistColdSpringHarborLaboratoryColdSpringHarbor,NewYork StevenH.Holtzman  M ChiefBusinessOfficerMillenniumPharmaceuticalsInc.Cambridge,Massachusetts BetteO.Kramer  R FoundingPresidentRichmondBioethicsConsortiumRichmond,Virginia BernardLo,M.D.  W DirectorPrograminMedicalEthicsUniversityofCaliforniaatSanFranciscoSanFrancisco,California LawrenceH.Miike,M.D.,J.D.  H!h] DirectorStateDepartmentofHealthHonolulu,Hawaii ThomasH.Murray,Ph.D.  0%P#b ProfessorandDirectorCenterforBiomedicalEthicsSchoolofMedicineCaseWesternReserveUniversityCleveland,Ohiò DianeScottJones,Ph.D.  *(i ProfessorDepartmentofPsychologyTempleUniversityPhiladelphia,Pennsylvania#)##X X1*# p0.p Ehd  (Xbb(̛   r XX   rNationalBioethicsAdvisoryCommissionStaff#r  z7##X X r[7#  P   ExecutiveDirector ,Acting D  WilliamF.Raub,Ph.D DeputyExecutiveDirector ,Acting   Ms.HenriettaD.HyattKnorr,M.A.̛̜*wddd Xdd Xdd X##w,dd ,z dd +   ` М ExecutiveAssistant  ( Ms.PatriciaNorris ProgramDirector   Ms.MargaretC.Quinlan % %К Analysts  ( Ms.EmilyFeinsteinMr.E.RandolphHull,Jr.Mr.SeanSimonMr.RobertTanner % % Director,HumanSubjectsProtectionsProject  l WilliamFreeman,M.D.,M.P.H. SeniorAnalyst (Volunteer) < JoelMangel,J.D. %, % SeniorSecretary  l Ms.RobinDorsey Secretary  <" Ms.LaShellGaskins %,# % %L$ % %L% % %4 &  % %4 ' % %!T(  % %!T) %М   ӀSeniorConsultant  <#* ЀKathiE.Hanna,Ph.D. .$,$+   .== %<#, % %%L -  %&%L .   &  ̜ `-'5   XfXXX Acknowledgments #X XXXf>#X XXX ԛ   >TheCommissionwishestoexpressitsspecialgratitudetothosescholarswhoonveryshortnoticeandunderverytighttimeconstraintspreparedattheCommissionsrequestverythoughtfulpresentationsandpapersontheissuesbeforetheCommission.Thesescholarsinclude:LoriB.Andrews,DanW.Brock,LisaCahill,CourtneyS.Campbell,RobertMullanCookDeegan,Ϝ!ElisaEiseman,!RabbiElliotDorff,NancyDuff,LeonR.Kass,BarthaMariaKnoppers,%%&&RuthMacklin,#G#$i$""#Gi#lbertC.Meilaender,FatherAlbertS.Moraczewski,JamesL.Nelson,))**StuartH.Orkin,''((JohnRobertson,JanetRossant,AbdulAzizSachedina,RabbiMosheTendler,andShirleyTilghman.TheCommissionalsowishestothankKathiE.Hanna,HenriettaHyattKnorr,andtheNBACstafffortheirunfailingcommitmentthroughtheninetydaysinwhichthisreportwasformulatedandproduced.OtherswhoprovidedsuperiorsupportincludeWilliamRaubinhisroleasActingExecutiveDirector,LilyEngstrom,HalThompson,DamonThompson,DonnaYoung,TimothyMorris,VitoOporto,JanetMiller,MarciaSnowden,andRosemarieMenz.̜#X XXX '?#  p Л  d  XfXXX  #7 XXdXXd7EXECUTIVESUMMARY #X XXXfD#X XXX   A- ) xdE0A  DTheideathathumansmightsomedaybecloned"createdfromasinglesomaticcellwithoutsexualreproduction"movedfurtherawayfromsciencefictionandclosertoagenuinescientificpossibilityonFebruary23,1997.Onthatdate,TheObserverbrokethenewsthatIanWilmut,a  Scottishscientist,andhiscolleaguesattheRoslinInstitutewereabouttoannouncethesuccessfulcloningofasheepbyanewtechniquewhichhadneverbeforebeenfullysuccessfulinmammals.Thetechniqueinvolvedtransplantingthegeneticmaterialofanadultsheep,apparentlyobtainedfromadifferentiatedsomaticcell,intoaneggfromwhichthenucleushadbeenremoved.Theresultingbirthofthesheep,namedDolly,onJuly5,1996,wasdifferentfrompriorattemptstocreateidenticaloffspringsinceDollycontainedthegeneticmaterialofonlyoneparent,andwas,therefore,a"delayed"genetictwinofasingleadultsheep.̜ F   Thiscloningtechniqueisanextensionofresearchthathadbeenongoingforover40yearsusingnucleiderivedfromnonhumanembryonicandfetalcells.Thedemonstrationthatnucleifromcellsderivedfromanadultanimalcouldbe"reprogrammed,"orthatthefullgeneticcomplementofsuchacellcouldbereactivatedwellintothechronologicallifeofthecell,iswhatsetstheresultsofthisexperimentapartfrompriorwork.Inthisreportthetechnique,firstϜdescribedbyWilmut,ofnucleartransplantationusingnucleiderivedfromsomaticcellsotherthanthoseofanembryoorfetusisreferredtoas somaticcellnucleartransfer.̜ F   WithindaysofthepublishedreportofDolly,PresidentClintoninstitutedabanonfederalfundingrelatedtoattemptstoclonehumanbeingsinthismanner.Inaddition,thePresidentaskedtherecentlyappointedNationalBioethicsAdvisoryCommission(NBAC)toaddresswithinninetydaystheethicalandlegalissuesthatsurroundthesubjectofcloninghumanbeings.Thisprovidedawelcomeopportunityforinitiatingathoughtfulanalysisofthemanydimensionsoftheissue,includingacarefulconsiderationofthepotentialrisksandbenefits.Italsopresentedanoccasiontoreviewthecurrentlegalstatusofcloningandthepotentialconstitutionalchallengesthatmightberaisedifnewlegislationwereenactedtorestrictthecreationofachildthroughsomaticcellnucleartransfercloning.̜ F   TheCommissionbeganitsdiscussionsfullyrecognizingthatanyeffortinhumanstotransferasomaticcellnucleusintoanenucleatedegginvolvesthecreationofanembryo,withtheapparentpotentialtobeimplantedinuteroanddevelopedtoterm.EthicalconcernssurroundingissuesofembryoresearchhaverecentlyreceivedextensiveanalysisanddeliberationintheUnitedStates.Indeed,federalfundingforhumanembryoresearchisseverelyrestricted,althoughtherearefewrestrictionsonhumanembryoresearchcarriedoutintheprivatesector.Thus,undercurrentlaw,theuseofsomaticcellnucleartransfertocreateanembryosolelyforresearchpurposesisalreadyrestrictedincasesinvolvingfederalfunds.Thereare,however,nocurrentfederalregulationsontheuseofprivatefundsforthispurpose.̜ F   Theuniqueprospect,vividlyraisedbyDolly,isthecreationofanewindividual +&* geneticallyidenticaltoanexisting(orpreviouslyexisting)person"a delayedgenetictwin.ϜThisprospecthasbeenthesourceoftheoverwhelmingpublicconcernaboutsuchcloning.Whilethecreationofembryosforresearchpurposesalonealwaysraisesseriousethicalquestions,theuseofsomaticcellnucleartransfertocreateembryosraisesnonewissuesinthisrespect.Theuniqueanddistinctiveethicalissuesraisedbytheuseofsomaticcellnucleartransfertocreatechildrenrelateto,forexample,serioussafetyconcerns,individuality,familyintegrity,andtreatingchildrenasobjects.Consequently,theCommissionfocuseditsattentionontheuseofsuchtechniquesforthepurposeofcreatinganembryowhichwouldthenbeimplantedinawoman'suterusandbroughttoterm.Italsoexpandeditsanalysisofthisparticularissuetoencompassactivitiesinboththepublicandprivatesector. F   Initsdeliberations,NBACreviewedthescientificdevelopmentswhichprecededtheRoslinannouncement,aswellasthoselikelytofollowinitspath.Italsoconsideredthemanymoralconcernsraisedbythepossibilitythatthistechniquecouldbeusedtoclonehumanbeings.Muchoftheinitialreactiontothispossibilitywasnegative.Carefulassessmentofthatresponserevealedfearsaboutharmstothechildrenwhomaybecreatedinthismanner,particularlypsychologicalharmsassociatedwithapossiblydiminishedsenseofindividualityandpersonalautonomy.Othersexpressedconcernaboutadegradationinthequalityofparentingandfamilylife. F   Inadditiontoconcernsaboutspecificharmstochildren,peoplehavefrequentlyexpressedfearsthatthewidespreadpracticeofsomaticcellnucleartransfercloningwouldundermineimportantsocialvaluesbyopeningthedoortoaformofeugenicsorbytemptingsometomanipulateothersasiftheywereobjectsinsteadofpersons.Arrayedagainsttheseconcernsareotherimportantsocialvalues,suchasprotectingthewidestpossiblesphereofpersonalchoice,particularlyinmatterspertainingtoprocreationandchildrearing,maintainingprivacyandthefreedomofscientificinquiry,andencouragingthepossibledevelopmentofnewbiomedicalbreakthroughs. F   Toarriveatitsrecommendationsconcerningtheuseofsomaticcellnucleartransfertechniquestocreatechildren,NBACalsoexaminedlong-standingreligioustraditionsthatguidemanycitizens'responsestonewtechnologiesandfoundthatreligiouspositionsonhumancloningarepluralisticintheirpremises,modesofargument,andconclusions.Somereligiousthinkersarguethattheuseofsomaticcellnucleartransfercloningtocreateachildwouldbeintrinsicallyimmoralandthuscouldneverbemorallyjustified.Otherreligiousthinkerscontendthathumancloningtocreateachildcouldbemorallyjustifiedundersomecircumstances,butholdthatitshouldbestrictlyregulatedinordertopreventabuses. F   ThepublicpoliciesrecommendedwithrespecttothecreationofachildusingsomaticcellnucleartransferreflecttheCommissionsbestjudgmentsaboutboththeethicsofattemptingsuchanexperimentand+our+,its,viewoftraditionsregardinglimitationsonindividualactionsinthenameofthecommongood.Atpresent,theuseofthistechniquetocreateachildwouldbea \+p&) prematureexperimentthatwouldexposethefetusandthedevelopingchildtounacceptablerisks.Thisinitselfmightbesufficienttojustifyaprohibitiononcloninghumanbeingsatthistime, +&* evenifsucheffortsweretobecharacterizedastheexerciseofafundamentalrighttoattempttoprocreate. F   Beyondtheissueofthesafetyoftheprocedure,however,NBACfoundthatconcernsrelatingtothepotentialpsychologicalharmstochildrenandeffectsonthemoral,religious,andculturalvaluesofsocietymeritedfurtherreflectionanddeliberation.Whetheruponsuchfurtherdeliberationournationwillconcludethattheuseofcloningtechniquestocreatechildrenshouldbeallowedorpermanentlybannedis,forthemoment,anopenquestion.Timeisanallyinthisregard,allowingfortheaccrualoffurtherdatafromanimalexperimentation,enablinganassessmentoftheprospectivesafetyandefficacyoftheprocedureinhumans,aswellasgrantingaperiodoffullernationaldebateonethicalandsocialconcerns.TheCommissionthereforeconcludedthatthereshouldbeimposedaperiodoftimeinwhichnoattemptismadetocreateachildusingsomaticcellnucleartransfer.$ !=  1          F   WithinthisoverallframeworktheCommissioncametothefollowingconclusionsandrecommendations:I.  TheCommissionconcludesthatatthistimeitismorallyunacceptableforanyoneinthe  publicorprivatesector,whetherinaresearchorclinicalsetting,toattempttocreateachildusingsomaticcellnucleartransfercloning.TheCommissionreachedaconsensusonthispointbecausecurrentscientificinformationindicatesthatthistechniqueisnotsafetouseinhumansatthispoint.Indeed,theCommissionbelievesitwouldviolateimportantethicalobligationswerecliniciansorresearcherstoattempttocreateachildusingtheseparticulartechnologies,whicharelikelytoinvolveunacceptableriskstothefetusand/orpotentialchild.Moreover,inadditiontosafetyconcerns,manyotherseriousethicalconcernshavebeenidentified,whichrequiremuchmorewidespreadandcarefulpublicdeliberationbeforethistechnologymaybeused. F   TheCommission,therefore,recommendsthefollowingforimmediateaction:"c  "cSj"0 F   0F^#F^#Acontinuationofthecurrentmoratoriumontheuseoffederalfundinginsupportofany  attempttocreateachildbysomaticcellnucleartransfer."cSjnj݌ ^#^# Ќ  "c  "ck"0 F   0F^#F^#Animmediaterequesttoallfirms,clinicians,investigators,andprofessionalsocietiesin l"  theprivateandnonfederallyfundedsectorstocomplyvoluntarilywiththeintentofthefederalmoratorium.Professionalandscientificsocietiesshouldmakeclearthatanyattempttocreateachildbysomaticcellnucleartransferandimplantationintoawoman'sbodywouldatthistimebeanirresponsible,unethical,andunprofessionalact."ckk݌ ^#^# Ќ  II.  TheCommissionfurtherrecommendsthat: '#& Ї"c  "cCn"0 F   0F^#F^#Federallegislationshouldbeenactedtoprohibitanyonefromattempting,whetherina  researchorclinicalsetting,tocreateachildthroughsomaticcellnucleartransfercloning.Itiscritical,however,thatsuchlegislationincludeasunsetclausetoensurethatCongresswillreviewtheissueafteraspecifiedtimeperiod(threetofiveyears)inordertodecidewhethertheprohibitioncontinuestobeneeded.Ifstatelegislationisenacted,itshouldalsocontainsuchasunsetprovision.Anysuchlegislationorassociatedregulationalsooughttorequirethatatsomepointpriortotheexpirationofthesunsetperiod,anappropriateoversightbodywillevaluateandreportonthecurrentstatusofsomaticcellnucleartransfertechnologyandontheethicalandsocialissuesthatitspotentialusetocreatehumanbeingswouldraiseinlightofpublicunderstandingsatthattime."cCn^n݌ ^#^# Ќ  III.  TheCommissionalsoconcludesthat: $  "c  "cr"0 F   0F^#F^#Anyregulatoryorlegislativeactionsundertakentoeffecttheforegoingprohibitionon   creatingachildbysomaticcellnucleartransfershouldbecarefullywrittensoasnottointerferewithotherimportantareasofscientificresearch.Inparticular,nonewregulationsarerequiredregardingthecloningofhumanDNAsequencesandcelllines,sinceneitheractivityraisesthescientificandethicalissuesthatarisefromtheattempttocreatechildrenthroughsomaticcellnucleartransfer,andthesefieldsofresearchhavealreadyprovidedimportantscientificandbiomedicaladvances.Likewise,researchoncloninganimalsbysomaticcellnucleartransferdoesnotraisetheissuesimplicatedinattemptingtousethistechniqueforhumancloning,anditscontinuationshouldonlybesubjecttoexistingregulationsregardingthehumaneuseofanimalsandreviewbyinstitution-basedanimalprotectioncommittees."crr݌ ^#^# Ќ  "c  "cw"0 F   0F^#F^#Ifalegislativebanisnotenacted,orifalegislativebaniseverlifted,clinicaluseof   somaticcellnucleartransfertechniquestocreateachildshouldbeprecededbyresearchtrialsthataregovernedbythetwinprotectionsofindependentreviewandinformedconsent,consistentwithexistingnormsofhumansubjectsprotection."cw w݌ ^#^# Ќ  "c  "cy"0 F   0F^#F^#TheUnitedStatesGovernmentshouldcooperatewithothernationsandinternational   organizationstoenforceanycommonaspectsoftheirrespectivepoliciesonthecloning"cyy݌ ^#^# Ќ  0 F 0F^#F^#ofhumanbeings. ^#^# IV.  TheCommissionalsoconcludesthatdifferentethicalandreligiousperspectivesand D$X" traditionsaredividedonmanyoftheimportantmoralissuesthatsurroundanyattempttocreateachildusingsomaticcellnucleartransfertechniques.Therefore,theCommissionrecommendsthat:"c  "c{"0 F   0F^#F^#Thefederalgovernment,andallinterestedandconcernedparties,encouragewidespread (#' andcontinuingdeliberationontheseissuesinordertofurtherourunderstandingoftheethicalandsocialimplicationsofthistechnologyandtoenablesocietytoproduceappropriatelong-termpoliciesregardingthistechnologyshouldthetimecomewhen +&* presentconcernsaboutsafetyhavebeenaddressed."c{{݌ ^#^# Ќ  V.  Finally,becausescientificknowledgeisessentialforallcitizenstoparticipateinafull  andinformedfashioninthegovernanceofourcomplexsociety,theCommissionrecommendsthat:"c  "c"0 F   0F^#F^#Federaldepartmentsandagenciesconcernedwithscienceshouldcooperateinseekingout t  andsupportingopportunitiestoprovideinformationandeducationtothepublicinthe"c#݌ ^#^# Ќ  0 F 0F^#F^#areaofgenetics,andonotherdevelopmentsinthebiomedicalsciences,especiallywheretheseaffectimportantculturalpractices,values,andbeliefs.#XXXX&D# 8 L ^#^# 7XXdXXd7  Xf9XXX (#  ChapterOne #XXXXf9#X&XXX@@*  XX&# ^ -#V^  ^ @$INTRODUCTION # ^ V^Ƃ#  ^#X&X脂#Ԉ  A- ) xdE0A  ̜Theideathathumansmightsomedaybecloned"createdfromasinglesomaticcellwithoutsexualreproduction"movedfurtherawayfromsciencefictionandclosertoagenuinescientificpossibilityonFebruary23,1997.Onthatdate,TheObserverbrokethenewsthatIanWilmut,a   Scottishscientist,andhiscolleaguesattheRoslinInstitutewereabouttoannouncethesuccessfulcloningofasheepbyanewtechnique.Thetechniqueinvolvedtransplantingthegeneticmaterialofanadultsheep,apparentlyobtainedfromadifferentiatedsomatic) !=  2      כcell,intoaneggfromwhich {   thenucleushadbeenremoved.Theresultingbirthofthesheep,namedDolly,onJuly5,1996appearstomarkyetanothermilestoneinourabilitytocontrol,refine,andamplifytheforcesofnature.̜ F   TheScottishsheepexperimentwasdifferentfrompriorattemptstocreateidenticaloffspringfromapairofadultanimals.Itusedacloningtechniquetoproduceananimalthatwasagenetictwinofanadultsheep.Putanotherway,Dollycontainedthegeneticmaterialofonlyoneparent.Thistechniqueoftransferringanucleusfromasomaticcellintoaneggisanextensionofresearchthathadbeenongoingforover40yearsusingnucleiderivedfromnonhumanembryonicandfetalcells.Thedemonstrationthatnucleifromcellsderivedfromanadultanimalcouldbe"reprogrammed,"orthatthefullgeneticcomplementofsuchacellcouldbereactivatedwellintothechronologicallifeofthecell,iswhatsetstheresultsofthisexperimentapartfrompriorwork.Inthisreportthetechnique,firstreportedbyWilmut,ofnucleartransplantationusingnucleiderivedfromsomaticcellsotherthanthoseofanembryoorfetusisreferredtoas somaticcellnucleartransfer. F   Forsometime,scientificevidencehassuggestedthatthegeneticmaterialcontainedindifferentiatedsomaticcellsmayretainthepotentialtodirectthedevelopmentofhealthyfertileadultanimals,butitscapacitytodosoremainedunproved(DiBernadino,1997).TheRoslinexperiment,therefore,wasasignificantscientificeventwithpotentiallyprofoundimplicationssinceitbringsusclosertothepossibilityofdevelopingacapacitytocreateclonehumanbeingsinanasexualmanner.Althoughforthepasttenyearsscientistshaveroutinelyclonedsheepandcowsfromembryocells,thiswasthefirstsuccessfulexperimentusingthenucleusofasomaticcellfromanadultanimaltocloneananimalthatmaturedtoafullydevelopedstate.̜ F   Theissuessurroundingthecloningofhumanbeingshavelongbeenthesubjectof periodicconcernanddebateamongphilosophers,scientists,ethicists,andothers,particularly _&s!% followingthepublicationofJoshuaLederbergs1966articleoncloningintheAmerican  Naturalist(Lederberg,1966).Nevertheless,theimpactofthesemostrecentdevelopmentson   ournationalpsychehasbeenquiteremarkable.Somecommentatorshavesuggestedthatthefurorarousedbythenewpossibilityforcloningisoutofproportiontomostoftheethical,legal,andmoralissuesitraises,sincethesesameissueshavebeenraisedbypreviousdevelopmentsandaresimplyemergingagaininanovelandstrikingform.Nevertheless,itisimportanttoacknowledgethatthepossibilitiesraisedbythisnewtechniquecertainlywouldbeunprecedentedandthatsomewouldconsideritsusetobeatrulyradicalstep.Thistypeofcloningwouldinvolvethreenoveldevelopments:thereplacementofsexualprocreationwithasexualreplicationofanexistingsetofgenes;theabilitytopredeterminethegenesofachild;andtheabilitytocreatemanygeneticallyidenticaloffspring.̜ F   Somescientistsweresurprisedthatthetechnicalbarriersofcelldifferentiationanddevelopmentseeminglycouldbesoeasilyovercomewhenusingsomaticcellsasthesourcefornucleartransfer.Thepublic"includingmanymembersofthescientificcommunity"respondedtoDollywithacombinationoffascination,hopeforusefulnewunderstandingsofhumanbiology,andprofoundconcern"evenalarm"abouttheprospectofbeingabletocreatewholehumansfromasinglesomaticcellvianucleartransfercloningtechniques.Althoughmuchoftheinitialpublicreactionwasoneoffear,concern,andseriousmoralreservationsaboutthepotentialuseorabuseofthisnewtechnologicalcapacity,afewvoiceswereheardcautiouslysuggestingthatabetterunderstandingofcelldynamicsinhumansandanimalsmightenableustodevelopnewcuresforvariousdiseases.Thus,itisimportanttoreflectnotonlyonthedangersandethicalreservationsbutalsoonthepotentialhumanbenefitsfromtheuseofthistypeofcloningthatmightariseinsuchareasastreatingparticularinfertilityproblems,transplantingcellsortissues,orpreventingcertaingeneticallytransmittedharmstooffspring. F   Afewoftheinitialobjectionstothisnewtypeofcloningwereeitherspeculativeorbasedonsimplemisunderstandings,suchas,thatcloningwouldallowfortheinstantaneouscreationofafullygrownadultfromthecellsofanindividual.Otherfearsstemmedfromtheincorrectideathatanexactcopy,althoughmuchyounger,ofanexistingpersoncouldbemade.Thisfearreflectsanerroneousbeliefthatone'sgenesbearasimplerelationshiptothephysicalandpsychologicaltraitsthatmakeupaperson.Althoughgenesprovidethebuildingblocksforeachindividual,itistheinteractionamongapersonsgeneticinheritance,thephysicalandculturalenvironment,andtheprocessoflearningthatresultintheuniquenessofeachindividualhuman.Thus,theideathatnucleartransplantationcloningcouldbeusedtore-createexemplaryorevilpeoplehasnoscientificbasisandissimplyfalse. F   Otherobjectionstonucleartransplantationcloning,however,arebasedoncarefullyarticulatedphilosophicalideals,deepculturalcommitments,orreligiousbeliefs,andthesedeservecontinuingandcarefulconsideration.Theseobjectionsreflectdeeplyheldbeliefsaboutthevalueofhumanindividualityandpersonalautonomy,themeaningoffamilyandthevalueofachild,respectforhumanlifeandthenaturalworld,andthepreservationoftheintegrityofthehumanspecies. +&* Ї F   ManypublicleadersintheUnitedStatesrespondedtotheannouncementaboutDollywithimmediateandstrongcondemnationofanyattempttoclonehumanbeingsinthisnewmanner.Thereasonsrangedfromfrighteningsciencefictionimagerytothejudgmentthatcloningofhumanbeingsisaseriousviolationofbasichumanrightsandhumandignity.Thereactionabroadwassimilar,withmanynationsseeminglyready"indirectlyordirectly"toprohibitcloninghumanbeingsinthisfashion.Indeed,manyinternationalorganizationssuchasUNESCOandtheCouncilofEuropehavealong-establishedandwell-articulatedconcernthatresearchandclinicalapplicationsinbiologyandgeneticsremainconsistentwithafundamentalcommitmenttohumandignityandhumanrights.Todate,atleastArgentina,Australia,GreatBritain,Denmark,Germany,andSpainhaveenactedlawsbanningcloninghumanbeings.Unfortunately,someofthedeepconcernssupportingsuchviewsandassociatedlegislationarestatedinvagueoroverlybroadterms.Thewidespreadpublicdiscomfort,evenrevulsion,aboutcloninghumanbeingsdeservesthebestarticulationpossible,ataskthattakestimeandrequirestheconsideredreflectionsofdiversegroupswithinAmericansocietyandabroad. F   Withindaysofthepublishedreportoftheapparentlysuccessfulcloningofasheepinthisnewmanner,PresidentClintoninstitutedabanonfederalfundingforresearchrelatedtocloningofhumanbeings.Inaddition,thePresidentaskedtherecentlyappointedNationalBioethicsAdvisoryCommission(NBAC)toaddresswithinninetydaystheethicalandlegalissuesthatsurroundthesubjectofcloninghumanbeings.Thisprovidedawelcomeopportunityforinitiatingathoughtfulanalysisofthemanydimensionsoftheissue,includingacarefulconsiderationofthepotentialrisksandbenefits.Italsopresentedanoccasiontoreviewthecurrentlegalstatusofcloningandthepotentialconstitutionalchallengesthatmightberaisedifnewlegislationwereenactedtorestrictthecreationofachildthroughsomaticcellnucleartransfer. F   TheCommissionbeganitsdiscussionsfullyrecognizingthatanyeffortinhumanstotransferasomaticcellnucleusintoanenucleatedegginvolvesthecreationofanembryo,withtheapparentpotentialtobeimplantedinuteroanddevelopedtoterm.Ethicalconcerns  surroundingissuesofembryoresearchhaverecentlyreceivedextensiveanalysisanddeliberationinourcountry.Indeed,federalfundingforhumanembryoresearchisseverelyrestricted,althoughtherearefewrestrictionsonhumanembryoresearchcarriedoutintheprivatesector.Thus,undercurrentlaw,theuseofsomaticcellnucleartransfertocreateanembryosolelyforresearchpurposesisalreadyrestrictedincasesinvolvingfederalfunds.Thereare,however,nocurrentregulationsontheuseofprivatefundsforthispurpose. F   Theuniqueprospect,vividlyraisedbyDolly,isthecreationofanewindividualgeneticallyidenticaltoanexisting(orpreviouslyexisting)person"a delayedgenetictwin.Thisprospecthasbeenthesourceoftheoverwhelmingpublicconcernaboutsuchcloning.Whilethecreationofembryosforresearchpurposesalonealwaysraisesseriousethicalquestions,theuseofsomaticcellnucleartransfertocreateembryosraisesnonewissuesinthis respect.Theuniqueanddistinctiveethicalissuesraisedbytheuseofsomaticcellnuclear *%) transfertocreatechildrenrelateto,forexample,serioussafetyconcerns,individuality,family integrity,andtreatingchildrenasobjects.Consequently,theCommissionfocuseditsattentionontheuseofsuchtechniquesforthepurposeofcreatinganembryowhichwouldthenbeimplantedinawoman'suterusandbroughttoterm.Italsoexpandeditsanalysisofthisparticularissuetoencompassactivitiesinboththepublicandprivatesector.  #XXXX&,#Xf9XXXControllingNature#XXXXf9#X&XXX  t  Y F   Humankind'seffortstocontrolnaturedatebackasfarasrecordedhistory.Inparticular,domesticatedplantsandanimalshavebeenthemainstayofouragriculturalheritage.Overtimehumanmasteryovernatureoftenhasbeenmet,quiteunderstandably,withoppositionandconcern,andfrequentlyhasbeenconsideredbysometobeanaffronttothenaturalorderofthingsorbyotherstobeatoddswithinterpretationsofGod'srevealedword.Indeedmanymythsandlegends,ancientaswellasmodern,dealdirectlywithhumankind'son-goingstruggletoensurethatthebenefitsofournewtechnologicalcapacitiesclearlyoutweightheharms"bothexpectedandunexpected.Theideathatourgrowingtechnologicalmasteryisfilledwithmoralambiguityandcapableofbothvastgoodandcatastrophicevilisdeeplyembeddedinmanyculturaltraditions. F   AprimeexampleisthemythologyoftheArgo,thefirstship,inclassicalGreekculture.TheGreeksseetheinitialactofshipbuildingasboththeoriginofcultureandtheoriginofdecline.Whilesailingenablesonetoencounterotherpersonsandotherpossibilities,italsobringsmaraudersandwar,anditsveryexistencebespeaksthedangerofunlimitedhumandesire.Thus,theabilitytobuildandsailboatsisbothaboonandacurse.Euripides'MedeastartswithalamentaboutthetreesthatwerecutdowntobuildtheArgoandtheothertroublesthatfollowed:8 F 0F"F"WouldthattheArgohadneverwingeditswaytothelandofColchis.... "" 8 F 0F"F"WouldthatpinetreeshadneverbeenfelledintheglensofMountPelionand "" 0 F 0F^#F^#furnishedoarsforthehandsoftheheroeswhoatPelias'commandsetforthin ^#^# 0 F 0F^#F^#questoftheGoldenFleece. ^#^#  F   Concernaboutourtoolsandtechnologyhasbeengreatlyacceleratedwiththecomingofmodernindustrializedsocieties.Isitpossible,somenowwonder,thatourconfidenceinhumancompetenceandtechnologymaybejustanothermyth?How,somearenowasking,canwefindsomemoralcompassormorallimittoourdesiretomastereverythingandpossessall?Onlysuchlimits,manywouldsay,cansaveusfromthemoralambiguityofourowncleverness. F   Inrecentyears,concernabouthumankind'scontrolovernaturehasbeenparticularlyacuteinrelationtothenewmoralchoicescreatedbythestunningdevelopmentsinthebiomedicalsciences,especiallyintheareaofhumanreproduction.Althoughpersonalreproductivehealthisconsideredtobe,inmostcases,aprivatematter,ongoingcontroversiesregardingthemoralstandingofhumangeneticmaterialandparticularhumaninterventionsinprocreationhavefocusedpublicattentionontheethicalandlegalimplicationsofnewreproductivetechniques.In +&* manycases,initialfearsgivewaytocautiousacceptance,butawarinesslingersthatiseasilyreawakenedwitheachnewadvance. F   Artificialinseminationbydonor,forexample,wasconsideredaformofadulterywhenfirstintroducedinthe1940s.Itisnowawidelyusedandacceptedpracticeinthetreatmentofinfertility,althoughsomecontinuetohaveseriousreservations.Whenprenataldiagnosiswasintroducedinthelate1960s,thepublicsimultaneouslywelcomedtheopportunitytopreventlethaldiseaseinnewbornsbutworriedabouttheuseofsuchtechniquestoselect"vanity"characteristicsornonmedicaltraitsinoffspring.ThebirthofLouiseBrown,conceivedviain L ` vitrofertilization,in1978wasanotherdramaticevent,providinganewandcontroversialmeans 8 L  toparenthood.Withallofthesetechnicaladvances,therehasbeenacontinuingdebateaboutsafety,legality,ethicalacceptability,andthegovernment'srighttointerveneinprivatematters. F   Researchitself,notjustitsclinicalapplication,hasoftensparkeddebate.Forexample,researchinvolvinghumanfetuseshasbeenasubjectofintensenationaldebateanddisagreementforovertwodecades(InstituteofMedicine,1994).Federalresearchinthisareacontinuestoberestrictedtothatwhichhaspotentialtherapeuticbenefittothefetus,orinvolvesnomorethanminimumrisktothefetusevenifpotentialbenefittothemothercanbedemonstrated.Restrictionsalsoremainregardingembryoresearch.DespitetherecommendationsoftheNationalInstitutesofHealthHumanEmbryoResearchPanel(1994),thatcertaintargetedandcarefullyregulatedresearchusingearlyhumanembryosbeeligibleforfederalfunds,inDecember1994thePresidentdirectedNIHnottoallocatefederalfundsforresearchprogramsthatinvolvedthecreationofhumanembryossolelyforresearchpurposes.ThisissuewasalsoaddressedbyCongress,whichinsertedlanguageintheFY96andFY97appropriationsbillsthatwidenedthepresidentialbantoprohibitvirtuallyallhumanembryoresearchconductedwithfederalfunds.WorkinthisareacontinuesintheUnitedStates,butitislargelylimitedtotheprivatesector,andthustakesplacewithoutanyfederalregulation. F   RecombinantDNAresearchrepresentsanotherexampleofcontroversyandintensedebate.Inthe1970s,concernsaboutthesafetyofunintendedreleaseofrecombinantorganismsledtoavoluntaryresearchmoratoriuminthescientificcommunityandthedevelopmentofguidelines(Fredrickson,1991).Similarly,allexperimentsinvolvinggenetherapy(treatmentofspecificdiseasesbyinsertinghumangenesintohumanpatients)aresubjecttoreviewandapprovalbyafederalbody. F   AssegmentsofhumanDNAorhumancellsbecamethefocusofstudyandtheobjectsofmanipulation,theiruseasresearchmaterialsraisedincreasinglyimportantethicalissuesabouthowthesematerialsareobtained,transformed,and,insomecases,usedtodevelopcommercialproducts(OfficeofTechnologyAssessment,1987).Suchresearchwithhumangeneticmaterialgeneratesquestionsaboutrespectforpersonsandthehumanbody,andthevalueandmoralstatustobeplacedoncellsandtissues.  *%)  F   Geneticandreproductivetechnologiesalsocauseconcernbecauseofthespecterof eugenicsandofrealorimaginedsocialcontrolthroughmanipulationofhumangenes.Geneticcontrolsuggestsbrokentaboos,and,inthewordsofHenryDavidThoreau,impliesthat menhavebecomethetoolsoftheirtools(Blank,1981).Whiletheseconcernsareoftensetagainstandpartlyattributabletoabackdropoffiction,fantasy,andmisunderstanding,theyare,moreimportantly,relatedtoprofoundconcernsregardingthenatureofhumankindanditsrelationshiptootheraspectsofthenaturalworld.* !=  3      ׀Whenthebizarreandfantasticscenariosareremoved,we t  areleftwithamyriadofreactions:sincereexpressionsofopposition;seriousmoralconcerns;newhopeforabetterunderstandingofhumanbiologyandtheprospectofcombatingcurrentlyuntreatableafflictions;callsformorestudy;andguardedstatementsabouttheneedforsomemeasureofcontrol(Macklin,1994;1997).  #XXXX&߮#Xf9XXXControllingScience#XXXXf9 #X&XXX     F   Withsomenotableexceptions,thescientificcommunityhasenjoyedforcenturiesagreatdealofautonomyindirectingandregulatingitsresearchagenda.Sincemidcentury,however,demandsforexternalregulationhaveincreased,inpartbecausemuchresearch,particularlyinthebiologicalsciences,ispubliclyfundedandthereforerequiressomeadditionalmeasureofaccountability.Moreimportantly,societyhasbecomemoresensitivetoconcernsaboutthedangers"particularlytohumanparticipants"oftheresearchitselfanditsfutureconsequences.Thus,ourevolvingmoralsensibilitiestogetherwiththespectacularadvancesinbiomedicalsciencehavegeneratednewethicalconcerns.AsBernardDavisofHarvardMedicalSchoolandothershavenoted,societysometimesseekstoregulateorrestrictresearchwhenitposesthespectersofdangerousorunfamiliarproducts,powers,orideas(Davis,1980).̜ F   Theregulationofsciencehasthusbecomepartofthelandscape,particularlyforthosewhoreceivefederalfunds(OfficeofTechnologyAssessment,1986).Inadditiontoenvironmental,health,occupational,andsafetyregulations,scientistsmustalsocomplywithanimalwelfareandhumansubjectsprotectionsandabidebyrestrictionsandmoratoriaonspecifictypesofresearch.Becausescienceisbothapublicandsocialenterpriseanditsapplicationcanhaveprofoundimpact,societyrecognizesthatthefreedomofscientificinquiryisnotanabsoluterightandscientistsareexpectedtoconducttheirresearchaccordingtowidelyheldethicalprinciples.Therearetimeswhenlimitsonscientificfreedommustbeimposed,evenifsuchlimitsareperceivedasanimpedimentbyanindividualscientist.Moreover,appropriateethicalconstraintsareamatterforbothscientistsandthebroaderpublictoformulateandimplement.Atthesametime,limitsonfreedomofinquirymustbejustified,andimpositionsonsuchfreedomshouldsatisfycertainconditions"forexample,thatthelimitsarenotarbitrary,that &0!$ theyemergefromthethoughtfulbalancingofcostsandbenefits,thattheyarenotunnecessarilyoppressive,thattheydonotlightlyimpingeonlongestablishedrightsandfreedoms,thatthereissomecontinuingpublicdiscoursewiththoseaffectedbytheban,andthatsuchlimitationsbeopentoreconsiderationinthelightofnewinformationandnewunderstanding.̛̜  #XXXX&`#Xf9XXXConsiderationofEthicalandReligiousPerspectives#XXXXf9#X&XXX    p̜ F   WhenthePresidentaskedNBACtotakeuptheissueofthecloningofhumanbeingsheadmonishedthat"anydiscoverythattouchesuponhumancreationisnotsimplyamatterofscientificinquiry,itisamatterofmoralityandspiritualityaswell."AlthoughwellawarethattheUnitedStatesConstitutionprohibitstheestablishmentofpoliciesthataresolelymotivatedby $8  religiousbeliefs,NBACsharedthePresidentsconcernandsoughtouttestimonyaboutthecloningofhumanbeingsfromleadingscholarsfromavarietyofreligioustraditions.InthesamespiritNBACalsocommissionedabackgroundpaperonthepositionsanumberofreligioustraditionshavetakenorareconsideringonthecloningofhumanbeings.̜ F   NBACfeltthiswasespeciallyimportantbecausereligioustraditionsinfluenceandshapethemoralviewsofmanyU.S.citizensandreligiousteachingsoverthecenturieshaveprovidedanimportantsourceofideasandinspiration.Althoughinapluralisticsocietyparticularreligiousviewscannotbedeterminativeforpublicpolicydecisionsthatbindeveryone,policymakersshouldunderstandandshowrespectfordiversemoralideasregardingtheacceptabilityofcloningofhumanbeingsinthisnewmanner.̜ F   Althoughsomereligiousresponsestothecloningofhumanbeingsthroughsomaticcellnucleartransferaretiedtightlytoparticularscripturaltextsorotherfaithcommitments,oftentheseideascanbestatedforcefullyintermsunderstandableandpersuasivetoallpersons,irrespectiveofspecificreligiousbeliefs.Forexample,appealmaybemadetoaviewofhumannatureorofhumanreason,ratherthanexclusivelytoareligioussourceofknowledgesuchasscriptureorrevelation.̜ F   NBACalsowantedtodeterminewhethervariousreligioustraditions,despitetheirdistinctivesourcesofauthorityandargumentation,reachsimilarconclusionsaboutthistypeofhumancloning.Aconvergenceofviewsacrossthesetraditions,aswellasacrossseculartraditions,wouldbeinstructive,evenifnotnecessarilydeterminative,forpublicpolicy.̜ F   WhilemanyAmericanslooktotheirreligiousfaithsformoralguidanceonissues,othersourcesofmoralknowledgeandinsightarealsoimportant.Manymoralconsiderationsthatwouldbewidelyacknowledgedaslegitimatedonotdependfortheirforceonparticularreligiouscommitmentsoraspecificphilosophicaloutlook.Forexample,theconvictionthatitiswrongtoharmachildisbroadlysharedamongAmericans.Ifyouinquirewhyitiswrongtoharmachild,peoplemaygivedifferentanswers.SomemayrefertotheirreligiousconvictionsthatachildisagiftfromGod.Othersmaysaythatitisalwayswrongtoharmaninnocentpersonwithoutsomecompellingreason.Tomanypeople,thisisabedrockprincipleofethics,evenifithasnosingle, +&* universallyacknowledgedfoundationinaspecificreligiousorphilosophicaltradition.Rather,itfindsitsfoundationinmanydifferentunderstandingsofmorality,somereligious,somesecular.MoralideassuchastheobligationnottoinflictharmonothersareaccessibletoallAmericansand,therefore,canprovidearobustfoundationforpublicpolicy.̜ F   Americahasavibranttraditionofethicaldialogueinwhichallareinvitedtoparticipate.Whatmoralconsiderationsdeserveourattentionandwhicharethemostimportantinrespondingtoaparticularissue?Thesearequestionsthatarisewitheverynewcontroversy.Whetherone'sethicalbeliefscomefromtheologicalcommitments,philosophicalarguments,orfromhard-wonlifeexperience,allvoicesshouldbewelcometotheconversation,andallthoughtfulviewsareentitledtoarespectfulhearing.WhiletoleranceisawidelyacceptedvirtueinAmericanitisimportanttoremindourselvesthatitisbuiltontheideaofmutualrespectandthecapacitytoaccept,wheneverpossible,themoralworthofotherswithwhomonemaydisagree.Tolerance,therefore,meansbothagreeingtodisagreeandacceptingthechallengeofsustainingacommunitywheremoralauthoritywill,tosomeextent,alwaysbecontested.̜ F   Policymakers,therefore,needtoconsiderarangeofmoralviewswhentheytrytodeterminewhetheraparticularpolicyisethicallyjustifiableaswellaspoliticallyfeasible.Aparticularpolicymaynotbepoliticallyfeasible,forinstance,ifitevokesthoughtful,widespreadandvigorousmoralopposition.Insuchcircumstancesitssocialcostsmayoutweighitsputativebenefits,andadditionaleducationanddeliberationmayberequiredbeforenewpoliciesareputinplace.#XXXX&#Xf9XXX @ ConsiderationofLawandPublicPolicy #XXXXf9#X&XXXԈ  4 ̜ F   Thepublicpolicychosenwithrespecttothecloningofhumanbeingsviasomaticcellnucleartransfershouldreflectakeenknowledgeofthescience,ourbestjudgmentsabouttheethicsofattemptingsuchanexperiment,andourtraditionsregardinglimitationsonindividualactionsinthenameofthecommongood.Americansinthisera,relativetoearliergenerations,haveawideinterestinandsubstantialknowledgeofscience.Nevertheless,intheweeksfollowingthereportofDolly,thepublic,themedia,andevensomescientistsdemonstratedasurprisinglackofunderstandingofthescienceinvolvedincloning.NBACbelievesthatpublicdebateaboutissuessuchashumancloningrequiresanevenmoreeducatedpopulace.Sciencepolicyhasbecomepublicpolicy,whichcanbedecidedwiselyonlybyaninformednation.̜ F   Americantraditionhasbeentoavoidprohibitingorregulatingpersonalactivities,absentacompellingreasonrelatedtoeffectsonothersorsocietyasawhole.Wheretheindividualactionsareexpressionsoffundamentalrights,suchastherighttofreespeechortherighttoprivacy,thereasonsforlimitationmustbecompelling,andthelimitationsmadeasminimalaspossible. F   Thepossibilityofcloninghumanbeingsinthisnewfashionappearstoraiseconcernsaboutdirectphysicalharmstothechildrenwhomayresult.Thisinitselfissufficienttojustifya +&* prohibitiononsuchattemptsatthistime,evenifsucheffortsweretobecharacterizedastheexerciseofafundamentalrighttoprocreate.Morespeculativepsychologicalharmstothechild,andeffectsonthemoral,religious,andculturalvaluesofsocietymaybeenoughtojustifyϜcontinuedprohibitionsinthefuture,butmoretimeisneededfordiscussionandevaluationoftheseconcerns. F   Initsdiscussionofpotentialpolicyoptions,NBACconsideredtherelativebenefitsofachievinganimmediateprohibitionthroughfederallegislationoncloninghumanbeingsusingsomaticcellnucleartransfertechniques.Italsoconsideredmoreindirectmeanstodetersuchexperiments. F   Indirect,non-legislativeoptionsconsideredbyNBACincludecooperationbytheprivatesector,bothresearchandclinical,inamoratoriumonsuchexperimentsand/orclinicalpractice,andthecontinuedprohibitionoftheuseoffederalfundstosupportsuchexperiments.TheAmericanMedicalAssociation,theWorldMedicalAssociation,andtheWorldHealthOrganization,forexample,havealreadycalledforsuchamoratoriumonclinicalactivities.̜ F   NBACalsoweighed,intermsofnucleartransplantationcloning,thepotentialimpactofapossiblelegislativemeasuretoextendbasichumansubjectsprotectionstoallresearchconductedintheUnitedStates.Thiswouldinsurethatanyresearcheffortstocloneahumaninthismannerwould,alongwithallotherresearchusinghumansubjects,becoveredbythetwinprotectionsofinformedconsentandappropriatescientificreviewtoinsureanethicallyacceptablebalancebetweenrisksandbenefits.Inlightoftheearlystateofanimalresearchinthisarea,suchprotectionsshouldpreventsuchcloningresearchfromgoingforwardatthistime.̜ F   Finally,NBACrecognizedthatcooperationwithothergovernmentsintheenforcementofanycommonelementsofourrespectivepoliciescouldstrengthenanyofthemeasuresadoptedbytheUnitedStates.Becausescienceisaglobalendeavor,internationalcooperationwouldensureconsistencyacrossbordersandenhancepublicconfidenceinscientificresearchgenerally.  #XXXX&#Xf9XXXProcessofNBACandOrganizationoftheReport#XXXXf9I#X&XXX     F   TheresultsofNBAC's90-dayanalysisarepresentedinthisreport.Initsdeliberations,NBACfocuseditsdiscussiononthescienceofthecloningofhumanbeingsusingthesomaticcellnucleartransfertechnique,andtheethical,religious,legal,andregulatoryimplicationsofcloninghumanbeingsinthismanner.ToaidinthesetasksNBACinvitedtestimonyfromanarrayofscientists,scientificsocieties,ethicists,theologians,andlegalexperts,andheardfromawidevarietyofinterestedpartiesduringthepubliccommentsessionateachmeeting.Inaddition,itcommissionednumerousbackgroundpapersfromrecognizedexpertstoinformitswork. F   Thisreportconsistsoffivechaptersinadditiontothisone.ChapterTwodescribesthescientificdevelopmentsthatprecededandmadepossiblethecloningofDollyandspeculatesonpotentialapplicationsofthisandrelatedtechnologies.ChapterThreepresentssomeofthekey +&* themesinreligiousinterpretationsandevaluationsofhumancloning.ChapterFouroutlinesthenumerousethicalconcernsraisedbytheprospectofcloninghumanbeingsviasomaticcellnucleartransfer.ChapterFivediscussesthelegalandpolicyissuesconsideredbytheNBACasitponderedvariousrecommendations.Thefinalsection,ChapterSix,presentstherecommendationsmadebyNBACinresponsetothePresidentsrequest.̜ F   Inmanyinstances,NBACfounditselfmovingatarapidpaceinonlypartlychartedwaters.Inthosetimesitreliedonitsindividualandcollectivewisdom,judgment,andmoralfoundations,andtheadviceofothers.NBACarguedanddebatedtheissuesasitsearchedforappropriateformulationsoftheproblemandforthewisdomtosuggestusefulpolicyoptions.WhilethemembersofNBAClearnedagreatdealduringitsdeliberations,wecouldnotreacharesolutiononalloftheissuesbeforeus.Nevertheless,itwasabletoaccomplishtwothings.First,itdevelopedasetofrecommendations,whicharesetoutinChapterSix.Second,itagreedthatitwasimportanttotakeanumberofstepstoensurethecontinuationofaninformednationaldiscussionoftheseissuesandotherdevelopmentsinthebiomedicalsciencesandclinicalpracticesthathaveanimpactonourmorallivesandculturaltraditions.    #XXXX&#Xf9XXXԛ  References #XXXXf9#X&XXX  ̜0 F  Blank,R.H.,ThePoliticalImplicationsofHumanGeneticTechnology(Boulder,CO:WestviewF^#F^#  F   Press,1981).̜0 F  Davis,B.D., Threespecters:Dangerousproducts,powers,orideas,inGeneticsandtheLawII, @0#  F^#F^#  F   A.MilunskyandG.J.Annas(eds.)(NewYork:PlenumPress,1980).̜0 F  DiBernadino,M.A.,GenomicPotentialofDifferentiatedCells(NewYork:ColumbiaUniversity @H# L `F^#F^#  F   Press,1997).̜0 F  Fredrickson,D.S., AsilomarandrecombinantDNA:Theendofthebeginning,inBiomedical (# $ F^#F^#  F   Politics,K.E.Hanna(ed.)(Washington,D.C.:NationalAcademyPress,1991).   ̜0 F  InstituteofMedicine,FetalResearchandApplications:AConferenceSummary(Washington, F^#F^#  F   D.C.:NationalAcademyPress,1994).̜0 F  Lederberg,J., Experimentalgeneticsandhumanevolution,TheAmericanNaturalist100:519F^#F^#  F   531,1966.̜0 F  Macklin,R., Splittingembryosontheslipperyslope:Ethicsandpublicpolicy,𛀜Kennedy\pF^#F^#  F   InstituteofEthicsJournal4(3)209225,1994. H\ ̜0 F  Macklin,R.,TestimonybeforetheNationalBioethicsAdvisoryCommittee,Washington,D.C., F^#F^#  F   March14,1997.̜0 F  NationalInstitutesofHealth,ReportoftheHumanEmbryoResearchPanel(Bethesda,MD:F^#F^#  F   NationalInstitutesofHealth,1994).̜0 F  OfficeofTechnologyAssessment,NewDevelopmentsinBiotechnology:OwnershipofHumanF^#F^#  F   TissuesandCells,OTABA337(Washington,D.C.:U.S.GovernmentPrintingOffice,    F   1987).̜0 F  OfficeofTechnologyAssessment,TheRegulatoryEnvironmentforScience(Washington,D.C.:X#l!F^#F^#  F   U.S.GovernmentPrintingOffice,1986).#XXXX&#  D$X"     ^ XXԛ7XXdXXd7V^  ^9(@ww%ChapterTwo# ^ V^#  ^@@*# ^ #V^  ^Ԉ  @P P THESCIENCEANDAPPLICATIONOFCLONING; !=  4      # ^ V^#  ^#X&X#   A- ) xdE0A  ThereportinFebruary1997thatscientistsinScotlandhadclonedasheep,Dolly,ledtomuch   publicdiscussionof"cloning"ofanimalsandspeculationaboutthepossibilityofcloninghumans.Theterm"cloning"isusedbyscientiststodescribemanydifferentprocessesthatinvolvemakingduplicatesofbiologicalmaterial.Inmostcasesisolatedgenesorcellsareduplicatedforscientificstudy,andnonewanimalresults.Thistypeofcloning,usinggenesandcells,hasledtomanymedicaladvancessuchasprovidinginsulintotreatdiabetesandtherapiesforhemophilia.Thesheepexperimentwasdifferent;itusedacloningtechniquecalled"somaticcellnucleartransfer"andresultedinananimalthatwasagenetictwin"althoughdelayedintime"ofanadultsheep.ThistechniqueoftransferringanucleusfromasomaticcellintoaneggthatproducedDollywasanextensionofexperimentsthathadbeenongoingforover40years.Theseexperimentswereaimedatunderstandinghowdevelopmentofananimalfromasinglefertilizedeggiscarriedout.Inrecentyearstheagriculturalindustryhasbeentryingtoimprovenucleartransplantationcloningtofacilitatethebreedingofdesirablelivestockandsomebiotechnologycompaniesareexploringwaystousenucleartransfercloningtoimprovetheproductionoftherapeuticdrugs.Inadditiontodrugproduction,understandingthedetailsofnucleartransplantationcloningmightleadtonewtherapiestotreathumandisease.Forinstanceitmightbepossibletogrowhumancellsandtissuesfortransplantationandgraftsthatwouldnotberejectedaftertransfer,astheyoftenaretoday.Thesekindsofbenefitsarecurrentlyonlyhypotheticalandmuchadditionalresearchwillbeneededinanimalsystems.AlthoughthebirthofDollywaslaudedasanamazingsuccess,infacttheprocedureisnotperfected.Onlyonesheepwasproducedfromovertwohundrednucleartransfers.Inaddition,itisnotyetclearwhetherDollyisnormalorwhethershecouldhavesubtleproblemsthatmightleadtoseriousdiseases.Usingthistechniquetoproduceahumanchildmightresultin,forexample,malformationsordiseaseduetoproblemsinherentinthetechnique.Thus,whileusinganimalstounderstandthebiologicalprocessthatproducedDollyholdsgreatpromiseforfuturemedicaladvances,thereisnocurrentscientificjustificationforattemptingtoproduceahumanchildatthistimewiththistechnique.   @*#XXXX&#Xf9XXXԈ  WhatisCloning? #XXXXf9#X&XXX "! Thewordcloneisusedinmanydifferentcontextsinbiologicalresearchbutinitsmostsimpleandstrictsense,itreferstoaprecisegeneticcopyofamolecule,cell,plant,animal,orhumanbeing.Insomeofthesecontexts,cloningreferstoestablishedtechnologiesthathavebeenpartof &!% agriculturalpracticeforaverylongtimeandcurrentlyformanimportantpartofthefoundationsofmodernbiologicalresearch. F   Indeed,geneticallyidenticalcopiesofwholeorganismsarecommonplaceintheplantbreedingworldandarecommonlyreferredtoas varietiesratherthanclones.Manyvaluablehorticulturaloragriculturalstrainsaremaintainedsolelybyvegetativepropagationfromanoriginalplant,reflectingtheeasewithwhichitispossibletoregenerateacompleteplantfromasmallcutting.Thedevelopmentalprocessinanimalsdoesnotusuallypermitcloningaseasilyasinplants.Manysimplerinvertebratespecies,however,suchascertainkindsofworms,arecapableofregeneratingawholeorganismfromasmallpiece,eventhoughthisisnotnecessarilytheirusualmodeofreproduction.Vertebrateshavelostthisabilityentirely,althoughregenerationofcertainlimbs,organs,ortissuescanoccurtovaryingdegreesinsomeanimals.̜ F   Althoughasingleadultvertebratecannotgenerateanotherwholeorganism,cloningofvertebratesdoesoccurinnature,inalimitedway,throughmultiplebirths,primarilywiththeformationofidenticaltwins.However,twinsoccurbychanceinhumansandothermammalswiththeseparationofasingleembryointohalvesatanearlystageofdevelopment.Theresultingoffspringaregeneticallyidentical,havingbeenderivedfromonezygote,whichresultedfromthefertilizationofoneeggbyonesperm.̜ F   Atthemolecularandcellularlevel,scientistshavebeencloninghumanandanimalcellsandgenesforseveraldecades.Thescientificjustificationforsuchcloningisthatitprovidesgreaterquantitiesofidenticalcellsorgenesforstudy;eachcellormoleculeisidenticaltotheothers.̜ F   Atthesimplestlevel,molecularbiologistsroutinelymakeclonesofdeoxyribonucleicacid(DNA),themolecularbasisofgenes.DNAfragmentscontaininggenesarecopiedandamplifiedinahostcell,usuallyabacterium.TheavailabilityoflargequantitiesofidenticalDNAmakespossiblemanyscientificexperiments.Thisprocess,oftencalledmolecularcloning,  isthemainstayofrecombinantDNAtechnologyandhasledtotheproductionofsuchimportantmedicinesasinsulintotreatdiabetes,tissueplasminogenactivator(tPA)todissolveclotsafteraheartattack,anderythropoietin(EPO)totreatanemiaassociatedwithdialysisforkidneydisease.̜ F   Anothertypeofcloningisconductedatthecellularlevel.Incellularcloningcopiesare X#l! madeofcellsderivedfromthesoma,orbody,bygrowingthesecellsincultureinalaboratory.Thegeneticmakeupoftheresultingclonedcells,calledacellline,isidenticaltothatoftheoriginalcell.This,too,isahighlyreliableprocedure,whichisalsousedtotestandsometimestoproducenewmedicinessuchasthoselistedabove.Sincemolecularandcellularcloningofthissortdoesnotinvolvegermcells(eggsorsperm),theclonedcellsarenotcapableofdevelopingintoababy.  )$( Л F   Thethirdtypeofcloningaimstoreproducegeneticallyidenticalanimals.Cloningofanimalscantypicallybedividedintotwodistinctprocesses,blastomereseparationandnuclear  transplantationcloning.   ̜ F   Inblastomereseparation,thedevelopingembryoissplitverysoonafterfertilizationwhenitiscomposedoftwotoeightcells(seefigure1).Eachcell,calledablastomere,isabletoproduceanewindividualorganism.Theseblastomeresareconsideredtobetotipotent,thatistheypossessthetotalpotentialtomakeanentireneworganism.Thistotipotencyallowsscientiststosplitanimalembryosintoseveralcellstoproducemultipleorganismsthataregeneticallyidentical.Thiscapabilityhastremendousrelevancetobreedingcattleandotherlivestock.̛ ^#^# ̜  m3<9)%`|0  `)ES  ~-m                  ^#^# F   Intheearly1980s,amoresophisticatedformofcloninganimalswasdeveloped,knownasnucleartransplantationcloning.Thenucleusofsomaticcellsisdiploid"thatis,itcontainstwosetsofgenes,onefromthemotherandonefromthefather.Germcells,however,containahaploidnucleus,withonlythematernalorpaternalgenes.Innucleartransplantationcloning,thenucleusisremovedfromaneggandreplacedwiththediploidnucleusofasomaticcell.Insuchnucleartransplantationcloningthereisasinglegenetic"parent,"unlikesexualreproductionwhereaneworganismisformedwhenthegeneticmaterialoftheeggandspermfuse(seefigureϜ2).Thefirstexperimentsofthistypeweresuccessfulonlywhenthedonorcellwasderivedfromanearlyembryo.Intheory,largenumbersofgeneticallyidenticalanimalscouldbeproducedthroughsuchnucleartransplantationcloning.Inpractice,thenucleifromembryoswhichhave  )$( developedbeyondacertainnumberofcellsseemtolosetheirtotipotency,limitingthenumberofanimalsthatcanbeproducedinagivenperiodoftimefromasingle,originatingembryo. 88^#^# 8888m3>9)%`|0, `(E, , 8 m 8888 8888 8888 8888 8888 8888 8888 8888 8888 8888 8888 8888 8888 ^#^#88̛̜ F   ThenewdevelopmentintheexperimentsthatWilmutandcolleaguescarriedouttoproduceDollywastheuseofmuchmoredevelopedsomaticcellsisolatedfromadultsheepasthesourceofthedonornuclei.Thisachievementofgestationandlivebirthofasheepusinganadultcelldonornucleuswasstunningevidencethatcelldifferentiationandspecializationarereversible.Giventhefactthatcellsdevelopanddivideafterfertilizationanddifferentiateintospecifictissue(e.g.,muscle,bone,neurons),thedevelopmentofaviableadultsheepfromadifferentiatedadultcellnucleusprovidedsurprisingevidencethatthepatternofgeneexpressioncanbereprogrammed.Untilthisexperimentmanybiologistsbelievedthatreactivationofthegeneticmaterialofmammaliansomaticcellswouldnotbecompleteenoughtoallowfortheproductionofaviableadultmammalfromnucleartransfercloning.  #XXXX&#Xf9XXXTheScienceThatLedtoDolly#XXXXf9G=#X&XXX      F   UntilthebirthofDolly,developmentalandmolecularbiologistsfocusedtheireffortsonunderstandingtheprocessesofcellulardifferentiation,theregulationofgenesduringthisprocess,thefactorsthatstimulatedifferentiation,andthereversibilityofthisprocess.Biologistshaveinvestigatedwhether,oncecellulardifferentiationoccurs,theprocessisreversible.ThesequestionshavebynomeansbeenfullyansweredbytheappearanceofDolly.Ifanything,theexistenceofDollystimulatesevenmorespeculationandinquiry.Thissectiondescribesthebackgroundofthesciencethatledtothebirthoftheclonedsheep,includingearlystudiesofdifferentiationanddevelopment,researchonregulationofgeneexpression,experimentsusingnucleartransferinanimals,andstudiesofcellprogramminganddivision.̜  +&* EarlyStudiesofDifferentiationandDevelopment   ̜ F   Nearlyeverycellcontainsaspheroidorganellecalledthenucleuswhichhousesnearlyallthegenesoftheorganism.GenesarecomposedofDNA,whichserveasasetofinstructionstothecelltoproduceparticularproteins.Althoughallsomaticcellscontainsthesamegenesinthenucleus,theparticulargenesthatareactivatedvarybythetypeofcell.Forexample,adifferentiatedsomaticcell,suchasaneuron,mustkeepasetofneural-specificgenesactiveandsilencethosegenesspecifictothedevelopmentandfunctioningofothertypesofcellssuchasmuscleorlivercells.̜ F   Investigationswhichbeganover40yearsagosoughttodeterminewhetheradifferentiatedsomaticcellstillcontainedallgenes,eventhoseitdidnotexpress.EarlyexperimentsinfrogsandtoadsbyGurdon(1962)andbyBriggsandKing(1952)providedstrongevidencethattheexpressionpotentialofthegenesindifferentiatedcellsisessentiallyunchangedfromthatoftheearlyembryo.Nucleifromdonordifferentiatedcellswereinjectedintorecipienteggsinwhichthenucleushadbeeninactivated(figure3).Thefirstseriesofexperimentsusedcellsfromtadpolesasthesourceofdonornuclei(Gurdon,1962)andadultfrogswereproduced,albeitataverylowefficiency.Althoughthecellsusedwerehighlyspecialized,theywerenotderivedfromtheadultfrog,sothecellsmightnothavebeenfullydifferentiated.m3@9)%`|0KRd `)E>ddd>m dd^#^# dddd dddd dddd dddd dddd dddd dddd dddd dddd dddd dddd dddd dddd dddd dddd dddd dddd dddd dddd ^#^#dd F   *%) Intheseexperiments,becauseisolatednucleiwereused,othercellularcomponentswerenot transferredtotherecipientegg.Amongthoseothercellularcomponentsisanorganellecalledthemitochondrion,theenergyproducingcomponentofthecell.Althoughmostofthegenesspecifyingthiscellularcomponentresideinthenucleus,themitochondrionitselfhousessomeofitsowngenes.Thus,insomaticcellnucleartransfer,mitochondrialgenesarenottransferredtotheenucleatedeggalongwiththenucleargenes.BecausetherearesomeseriousdiseasesϜassociatedwithmitochondrialgenes,nucleartransplantationcouldallowanembryotodevelopwithnew,healthymitochondriafromadonor.̜ F   Gurdonandcolleaguesperformedanothercarefullycontrolledseriesofexperimentsinwhichtheyusednucleifromadultfrogskincellsfortransfertoanenucleatedegg(Gurdon,etal.,1975).Fourpercentofthenucleitransferredeventuallygaverisetofullydevelopedtadpoles.Theseexperimentsprovidedevidencethatthegenescontainedinthenucleiofdifferentiatedcellscouldbereactivatedbythecytoplasmoftheeggandthusdirectnormaldevelopment,butonlyuptoacertainstage.Noviableadultfrogeverdevelopedfromthesetadpolesandtherewasadecreaseinthenumberoftadpolesbornastheageofthetransferrednucleusincreased.Thisleftopenthepossibilitythatcompletereactivationoftheadultnucleuswaspreventedbysomeirreversiblechangeinthegeneticmaterial,andthattherewasaprogressivedeclineinnuclearpotentialwithage. F   Carefulanalysis,however,suggestedthatthemajorreasonfordevelopmentalfailureofthetransplantedembryosappearedtobechromosomalabnormalitiesthatoccurredduringtheprocessofnucleartransplantationitself.Therateofcelldivisionofadultcellsismuchslowerthanthatofthecellsoftheearlyfrogembryo.Thus,inreality,forthistechniquetoworkitwouldbenecessarythatthetransplantedadultnucleusreprogramitsgeneexpression,replicateitsDNA,andenterthenormalembryoniccelldivisioncyclewithinanhourofnucleartransfer.Itisremarkable,giventhemechanicsandtimingoftheprocess,thatanynucleifromadultsomatic  cellsweresuccessfulingeneratinganembryo.Althoughtheydidnotproducenormaladultanimals,theamphibiannucleartransferexperimentsofGurdonandotherssucceededindemonstratingthatthedifferentiatedstateofadultsomaticcellsdonotinvolvemajorirreversiblechangesintheirDNA. RegulationofGeneExpression  l"  ̜ F   Inrecentyears,ithasbeendeterminedthatmostpatternsofdifferentiatedgeneexpressionaremaintainedbyactivecontrolmechanisms,inwhichparticulargenesareturnedonoroffbyregulatoryproteins(Blau,1992).Furtherstudiessuggestedthatitmightbepossibletoreprogramthegeneexpressionofsomaticcellssothattheyperformadifferenttask.Theroleofaparticularcelltype(e.g.,muscle,liver,orskin)dependsonthecombinationofregulatoryproteinsitexpresses.Whileincertainspecializedcells,suchaswhitebloodcells,actualrearrangementsanddeletionsofDNAoccur,forthemostpart,however,geneexpressionisnotregulatedbytheloss ofDNAbutbytheturningoffofspecificgenes.Thus,itshouldbepossibletoactivateor *%) inactivatealmostanygeneinacell,giventherightcellularenvironmentcontainingtheappropriateregulatorymolecules. ̜ F   Toreprogramthegeneexpressionofasomaticcellitisnotessentialtofuseitwithanegg;insomecasesre-programmingcanoccurthroughfusionoftwoadultcells.Cellfusionexperiments,inwhichdifferentsomaticcelltypesarefused,havedemonstratedthatextensivereprogrammingofdifferentiatednucleicanoccur.Forexample,whenmusclecellsarefusedwithnon-musclecellsofvarioussorts,muscle-specificgenesareactivatedinthenon-musclecells(Blauetal.,1985),and,similarly,genesthatcodeforhemoglobincanbeactivatedinmanycelltypesafterfusionwithredbloodcells(BaronandManiatis,1986).Theseandotherkindsofexperimentshaveledtotheisolationofspecificfactorsthatregulatecelldifferentiation,suchasthegenethatregulatestheformationofmusclecells(Weintraub,1993).̜ F   Thesestudieshavefurtherdemonstratedthatthestabilityofthedifferentiatedstateisnotabsolute.Thus,giventheappropriateregulatorymoleculesandenoughtimetoreprogramanadultnucleus,somaticcellscanre-initiateearlierprogramsofdifferentiation. NuclearTransferinMammals   ̜ F   Experimentsinmammalshavealsosuggestedthatispossibletoreprogramadultsomaticcells.Followingsuccessinthenucleartransferexperimentsinfrogs,scientistsattemptedtorepeattheexperimentsinmice.Itwasknownthatearlydevelopmentoccursataconsiderablyslowerrateinmammalsthanamphibians,givinghopethatreprogrammingofthedonornucleuswouldoccurmoreefficiently.Forexample,thefirstcelldivisioninmiceoccursaboutadayafterfertilization,givingampletime,itwasthought,forthereprogrammingofgeneexpressionandadjustmentofthecelldivisioncycle.Thisprovednottobethecase.Earlyexperimentsshowedthatnucleifromsomaticcellsfusedwithfertilizedeggsdidnotundergonucleardivision(Graham,1969). F   However,aseriesofexperimentsinmiceinthemid1980sshowedthatnucleicouldbesuccessfullyexchangedbetweenfertilizedeggs,with90percentreachingtheblastocyststageofembryonicdevelopmentandbeyond(McGrathandSolter,1984).Nucleirecoveredandtransplantedfromembryosatthetwo-cellstagecoulddirectdevelopmenttotheblastocyststage.Nucleitransferredfromembryosatlaterstages,however,couldnotsuccessfullyrecapitulatedevelopment.Infact,inmice,nucleishowlesstotipotencythanwholecells.Manyexperimentshaveshownthatblastomeresuptotheearlyblastocyststagearestilltotipotentwhencombinedwithotherembryoniccells(RossantandPedersen,1986).Thismeansthatthefailureofnuclearreprogramminghastobetheresultofsomethingotherthanirreversiblechangestothegeneticmaterialofthecells.In1986,Willadsenreportedexperimentswithsheep.Unlikethesituationinmice,enucleatedeggsfromsheepcouldbefusedwithblastomerestakenfromembryosattheeight-cellstagetoprovidedonornucleiandviableoffspringwereproduced(Willadsen,1986). +&* Мm3B9)%`|04l `)E4l4lhVmߛ hh^#^# hhhh hhhh hhhh hhhh hhhh hhhh hhhh hhhh hhhh hhhh hhhh hhhh hhhh hhhh̜ hhhh ^#^#hh̛ F   Recentexperimentshaveusednucleartransferintoenucleatedunfertilizedeggs(figure4).Usingtheseveryearlystageeggsprolongstheperiodofpossiblereprogrammingbeforethedonornucleushastoundergothefirstdivision.Andtheadventinthelastfewyearsofelectrofusionforbothfusionofcellsandactivationoftheegghasbeenanothermajoradvance,becauseactivationandfusionoccursimultaneously.Becausetheseexperimentsusefusionoftwocellsandnotsimpleinjectionofanisolatednucleus,allofthecellularcomponentsaretransferred.Thus,themitochondria,whichcontainsomegenesoftheirown,aretransferredalongwiththenucleus.Becauseanenucleatedeggalsocontainsmitochondria,theresultofafusionexperimentisacellwithamixtureofmitochondriafromboththedonorandtherecipient.Sincethemitochondrialgenesrepresentanextremelysmallproportionofthetotalnumberofmammaliangenes,mixingofmitochondriaperseisnotexpectedtohaveanymajoreffectsonthecell.However,iftheϜnucleusdonorsuffersfromamitochondrialdisease,andtheeggdonordoesnot,thenmixtureofthemitochondriamaysignificantlyalleviatethedisease.̜ F   Overthepasttenyearsorsotherehavebeennumerousreportsofsuccessfulnucleartransferexperimentsinmammals,nearlyallofthemusingcellstakendirectlyfromearlyembryos.Theoldestembryonicnucleusthatcansuccessfullysupportdevelopmentdiffersamongspecies.Four-cellblastomerenucleihavebeensuccessfullyusedinpigs(Prather,etal.,1989).Inmice,nonucleusolderthantheeight-cellstagehasbeenusedsuccessfully(Cheong,etal.,1993).Inrabbits,32-to64-cellearlyembryoscanbeusedasnucleardonors(Yang,etal.,1992).Incowsandsheep,cellsfromwhatiscalledtheinnercellmass(ICM)ofthe120-cellblastocyststage(seefigure1)havebeenusedsuccessfully(CollasandBarnes,1994;SmithandWilmut,1989).Indeed,inbothcowsandsheep,celllineshavebeenmadefromtheseICMcellsandnucleifromthesecellshavebeenusedtoreprogramdevelopmentaftertransferintoenucleatedunfertilizedeggs. +&* М F   InthefirstexperimentsofthissortbySimsandFirst(1994),cowcellsderivedfromembryosweregrowninthelaboratoryforupto28days,andthenusedasnucleardonors,withoutanyattemptatsynchronizationofthecelldivisioncycleofthedonorcells.Ofthosesuccessfullyfusedwitheggs,24percentdevelopedtotheblastocyststage,and4/34(12percent)oftheblastocyststransferredtorecipientcowsdevelopedintonormalcalves.Thissuccessratecomparesfavorablywiththoseseenusingearlierblastomeresandsuggeststhatitmightbepossibletoachievesuccessfulnucleartransferfrompermanentcelllinesestablishedfromearlyembryos. ReprogrammingofNucleiandSynchronizationoftheCellDivisionCycle  8 L  ̜ F   Therehasbeensomestudyoftheeventsthatoccuronceatransferrednucleusisexposedtothecytoplasmoftheeggandsome,butnotall,oftheparametersthataffectsuccessofnucleartransferareknown(Fulka,etal.,1996).Enucleatedeggsusedforfusiononlyproceedtodivisionafteractivationbysomeartificialsignal,suchastheelectricalcurrentusedintheelectrofusiontechnique.Whendonornucleiareintroducedintotheenucleatedegg,theyusuallyundergoDNAreplication,nuclearenvelopebreakdown,andchromosomecondensation.Afteractivationoftheeggthenuclearenvelopeisreformedaroundthedonorchromosomes.Thenucleusnowtakesontheappearanceofatypicaleggnucleusatthisstage,whichislargeandswollen.Itisassumedthatthisprocessbeginsthereprogrammingofthetransferreddonornucleusbyexposingthechromosomestotheeggcytoplasmandbeginningtheexchangeofegg-derivedproteinsforthedonornucleusownproteins(PratherandFirst,1990).̜ F   Itisnotclearwhetherexposuretoproteinsfoundintheearlieststagesofdevelopmentand/ornuclearswellingisaprerequisiteforreprogrammingforlaterdevelopment.Experimentsinanumberofspecieshaveshownthat,whennucleiarefusedwitheggsthathavebeenactivatedsomehourspriortofusion,noDNAreplication,chromosomecondensation,ornuclearswellingoccurs,butnormaldevelopmentcantranspire(Campbell,etal.,1994;Stice,etal.,1994). F   Onceagain,itisnotobviouswhichoftheprocessesdescribedabovearerequiredforϜnormaldevelopment.Inrabbits,cows,sheep,andmice(Cheong,etal.,1993;Collas,etal.,1992)experimentshaveshownthatnucleifromcellsintheearlyphasesofthecelldivisioncycledobetterthancellsinlaterstages.Inthefirstphaseofthecellcycle,termedG1(forGapphase1),cellscontainonlyonecompletesetofchromosomesandarerelativelyquiescent.TheythenenteraperiodofDNAsynthesisorreplication,calledS-phase,followedbyarestphase,calledG2(Gapphase2),atwhichtimetheyeachhaveaduplicatecopyofeachchromosome.Thisdoublingofthechromosomesisinpreparationforcelldivisionwhereanequalnumberwillbedividedbetweenthetwodaughtercells.BecauseDNAreplicationisinducedafternucleartransfer,any '"% nucleusthathasinitiatedreplicationbeforetransferwillendupwithtoomuchDNA,whichwill '#& likelyresultinchromosomeanomalies.Thus,theneedtotransfernucleiintheG1phasebeforereplicationisinitiated,islikelytobeimportanttoavoidchromosomedamagethatwillpreventdevelopmentoftheembryointoaviableoffspring. +&*  ChangesinTechniquethatAllowedfortheBirthofDolly    F   InbackgroundworkthatprecededthebirthofDolly,Wilmutandcolleaguesestablishedcelllinesfromsheepearlyembryos,orblastocysts,andusedthesecellsasnucleardonors(Campbell,etal.,1996).Inanattempttoavoidtheproblemsofnucleartransferofnon-G1nucleiintoactivatedeggs,theystarvedthedonorcelllinebyremovingallnutrientsfromthemediumpriortonucleartransfer.Underthesestarvationconditions,thecellsexitthecellcycleandentertheso-called G0state(Gapphase0),similartotheG1phaseinwhichchromosomeshavenotreplicated.FusionofG0nucleiwitheggsensuresthatthedonorchromosomeshavenotinitiatedreplicationpriortofusion.ItwasalsosuggestedthattheG0statemightactuallyincreasethecapacityofthenucleustobereprogrammedbytheeggcytoplasm.However,thereiscurrentlynodirectevidencetosupportthis,nortoconcludethatnucleisynchronizedintheG0stageareanybetterthannucleisynchronizedinG1.ForWilmutandcolleagues,approximately14percentoffusionsresultedindevelopmentofblastocysts,and4/34(12percent)embryostransferreddevelopedintolivelambs.Twodiedshortlyafterbirth.Thesuccessrateinsheepandcowexperimentswasalmostidentical,andsuggeststhatdivisionofcellsincultureformanydaysdoesnotinhibittheabilityoftheirnucleitobereprogrammedbytheeggenvironment.Couldthesamebetrueofnucleifromfullydifferentiatedsomaticcells?̜ F   AllofthisbackgroundworkleduptoDolly(Wilmut,etal.,1997).Wilmutandcolleaguestooklateembryo,fetalcellcultures,andcellculturesderivedfromthemammaryglandofanadultsheepandappliedthesameapproachofsynchronizingthecellsintheG0stagepriortonucleartransfer.Theyreportedsuccessfulproductionofliveoffspringfromallthreecelltypes,althoughonly29of277(11percent)ofsuccessfulfusionsbetweenadultmammaryglandnucleiandenucleatedoocytesdevelopedtotheblastocyststage,andonly1of29(3percent)blastocyststransferreddevelopedintoalivelamb.Thisexperimentwas,infact,thefirsttimeanyfullydevelopedanimalhadbeenbornfollowingtransferofasomaticcellnucleus,sincetheearlierfrogexperimentsonlygeneratedtadpoles. F Itshouldbenoted,however,thattheamountofnewinformationregardingthestabilityofthedifferentiatedstatederivedfromthisexperimentissmall,asnoattemptwasmadetodocumentthatthedonorcellswerefullydifferentiatedcells,thegenesofwhichexpressedspecializedmammaryglandproteins.Intheearlierexperimentswithfrogs,thefactthatthedonorcellswerefullydifferentiatedwasdocumentedinsuchamanner.Inthepresentcase,Dollycouldhavebeenderivedfromaless-differentiatedcellinthepopulation,suchasamammarystemcell. @ #XXXX&=#Xf9XXXRemainingScientificUncertainties#XXXXf9#X&XXX  &0!$ ̜ F   Severalimportantquestionsremainunansweredaboutthefeasibilityinmammalsofnucleartransfercloningusingadultcellsasthesourceofnuclei:  )$( М F   First,cantheprocedurethatproducedDollybecarriedoutsuccessfullyinothercases?Onlyoneanimalhasbeenproducedtodate.Thus,itisnotclearthatthistechniqueisreproducibleeveninsheep. ̜ F   Second,aretheretruespeciesdifferencesintheabilitytoachievesuccessfulnucleartransfer?Ithasbeenshownthatnucleartransferinmiceismuchlesssuccessfulthaninlarger   domesticanimals.Partofthisdifferencemayreflecttheintensityofresearchinthisareainthelasttenyears;agriculturalinterestshavemeantthatmorenucleartransferworkhasbeenperformedindomesticanimalsthaninmice.Butpartofthespeciesdifferencesmayberealandnotsimplyreflectthegreaterrecenteffortinlivestock.Forexample,inorderforadifferentiatednucleustoredirectdevelopmentintheenvironmentoftheegg,itsconstellationofregulatoryproteinsmustbereplacedbythoseoftheeggintimefortheembryotousethedonornucleustodirectnormaldevelopmentoftheembryo.Thespeciesdifferencemaybetheresultofthedifferenttimesofembryonicgeneactivation.̜ F   Inmammals,unlikemanyotherspecies,theearlyembryorapidlyactivatesitsgenesandcannotsurviveonthecomponentsstoredintheegg.Thetimeatwhichembryonicgeneactivationoccursvariesbetweenspecies"thelate2-cellstageinmice(Schultz,1993),the4-8cellstageinhumans(Braude,etal.,1988)andthe8-16cellstageinsheep.Thelateronsetofembryonicgeneactivationandtranscriptioninsheepprovidesanadditionalroundortwoofcelldivisionsduringwhichnuclearreprogrammingcanoccur,unliketherapidgenomeactivationinthemouse.Furthercross-speciescomparisonsareneededtoassesstheimportanceofthisdifferenceinthetimeofgenomeactivationforthesuccessofnucleartransferexperiments.Inhumans,forexample,thetimeperiodbeforegeneactivationisveryshort,whichmightnotpermittheproperreprogrammingofgenesafternucleartransfertoallowforsubsequentnormaldevelopment.̜ F   Third,willthephenomenonofgeneticimprintingaffecttheabilityofnucleifromlaterstagestoreprogramdevelopment?Inmammalsimprintingreferstothefactthatthegenesinheritedonthechromosomesfromthefather(paternalgenes)andthosefromthemother(maternalgenes)arenotequivalentintheireffectsonthedevelopingembryo(Solter,1988).Someheritableimprintisestablishedonthechromosomesduringthedevelopmentoftheeggandthespermsuchthatcertaingenesareexpressedonlywheninheritedfromthefatherormother.Imprintingexplainswhyparthenogeneticembryos,withonlymaternallyinheritedgenes,andandrogeneticembryos,withonlypaternallyinheritedgenes,failtocompletedevelopment(FundeleandSurani,1994).Nucleitransferredfromdiploidcells,whetherembryonicoradult,shouldcontainmaternalandpaternalcopiesofthegenome,andthusnothaveanimbalance &0!$ betweenthematernallyandpaternallyderivedgenes.̜ F   Thesuccessfulgenerationofanadultsheepfromasomaticcellnucleussuggeststhattheimprintcanbestable,butitispossiblethatsomeinstabilityoftheimprint,particularlyincellsin culture,couldlimittheefficiencyofnucleartransferfromsomaticcells.Itisknownthat *%) disturbancesinimprintingleadtogrowthabnormalitiesinmiceandareassociatedwithcancerandraregeneticconditionsinchildren.  F   Fourth,willcellularagingaffecttheabilityofsomaticcellnucleitoprogramnormaldevelopment?Assomaticcellsdividetheyprogressivelyageandthereisnormallyadefinednumberofcelldivisionsthattheycanundergobeforesenescence.Partofthisagingprocessinvolvestheprogressiveshorteningoftheendsofthechromosomes,thetelomeres,andothergeneticchanges.Germcells(eggsandsperm)evadetelomereshorteningbyexpressinganenzyme,telomerase,thatcankeeptelomeresfulllength.Itseemslikelythatreturninganadultmammaliannucleustotheeggenvironmentwillexposeittosufficienttelomeraseactivitytoresettelomerelength,sinceoocyteshavebeenfoundtobepotentsourcesoftelomeraseactivity(MantellandGreider,1994).̜ F   Fifth,willthemutationsthataccumulateinsomaticcellsaffectnucleartransferefficiencyandleadtocancerandotherdiseasesintheoffspring?Ascellsdivideandorganismsage,   mistakesandalterations(mutations)intheDNAwillinevitablyoccurandwillaccumulatewithtime.Ifthesemistakesoccurinthespermortheegg,themutationwillbeinheritedintheoffspring.Normallymutationsthatoccurinsomaticcellsaffectonlythatcellanditsdescendantswhichareultimatelydispensable.Nevertheless,suchmutationsarenotnecessarilyharmless.Sporadicsomaticmutationsinavarietyofgenescanpredisposeacelltobecomecancerous.Transferofanucleusfromasomaticcellcarryingsuchamutationintoaneggwouldtransformasporadicsomaticmutationintoagermlinemutationthatistransmittedtoallofthecellsofthebody.Ifthismutationwerepresentinallcellsmayleadtoageneticdiseaseorcancer.Therisksofsucheventsoccurringfollowingnucleartransferaredifficulttoestimate.  #XXXX&#Xf9XXXWhyPursueAnimalCloningResearch?#XXXXf9#X&XXX      ̜ F   Researchonnucleartransfercloninginanimalsmayprovideinformationthatwillbeusefulinbiotechnology,medicine,andbasicscience.Someoftheimmediategoalsofthisresearchare:"c  "c"0 F   togenerategroupsofgeneticallyidenticalanimalsforresearchpurposes"c݌!F^#F^# Ќ  "c  "cZ"0 F   torapidlypropagatedesirableanimalstocks"cZu݌l" F^#F^# Ќ  "c  "c"0 F   toimprovetheefficiencyofgeneratingandpropagatingtransgeniclivestock"c-݌X#l!F^#F^# Ќ  "c  "c"0 F   toproducetargetedgeneticalterationsindomesticanimals"c݌D$X"F^#F^# Ќ  "c  "c"0 F   topursuebasicknowledgeaboutcelldifferentiation"c̦݌0%D #F^#F^# Ќ  CloningAnimalsforResearchPurposes  '"% ̜ F   Inbredstrainsofmicehavebeenamainstayofbiologicalresearchforyearsbecausetheyareessentiallygeneticallyidenticalandhomozygous(i.e.,bothcopiesofeachgeneinheritedfromthemotherandfatherareidentical).Experimentalanalysisissimplifiedbecausedifferencesingeneticbackgroundthatoftenleadtoexperimentalvariationareeliminated.Generatingsuch +&* homozygousinbredlinesinlargeranimalsisdifficultandtimeconsumingbecauseofthelonggestationtimesandsmallnumbersofoffspring.Theconceptofgeneratingsmallgroupsofidenticalanimalsbynucleartransferhasbeenproposedasanalternativestrategytoobtainingageneticallyidenticalgroupofanimals,andapparentlyunderliesarecentreportfromOregononsuccessfulnucleartransferfromearlyembryonicnucleiinrhesusmacaquemonkeys(WeissandSchwartz,1997).̜ F   Repeatedcyclesofnucleartransfercanexpandthenumberofindividualanimalsderivedfromonedonornucleus,allowingmoreidenticalanimalstobegenerated.Thefirstnucleartransferembryoisallowedtodividetoearlyblastomerestagesandthenthosecellsareusedasdonornucleiforanotherseriesoftransfers.Thisprocesscanbecarriedonindefinitely,intheory,althoughpracticesuggeststhatsuccessfulfusionratesdeclinewitheachcycleoftransfer.Oneexperimentincows,forexample,produced54earlyembryosafterthreecyclesoftransferfromasingleblastomerenucleusfromoneinitialembryo(SticeandKeefer,1993).Viablecalveswereproducedfromallthreecyclesofnucleartransfer.̀ F   Thisapproachislikelytobelimitedinitsusefulness,however.Agroupofcloned  animalsderivedfromnucleartransferfromanindividualanimalisself-limited.Unlesstheyarederivedfromaninbredstockinitially,eachclonederivedfromoneindividualwilldiffergeneticallyfromaclonederivedfromanotherindividual.Onceaclonedanimalismatedtoproduceoffspring,theoffspringwillnolongerbeidenticalduetothenaturalprocesseswhichshuffleorrecombinegenesduringdevelopmentofeggsandsperm.Thuseachmemberofaclonehastobemadeforeachexperimentbynucleartransfer,andgenerationofalargeenoughnumberofclonedanimalstobeusefulasexperimentalgroupsislikelytobeprohibitivelyexpensiveinmostanimals.̜ AdvantagesofNuclearTransferCloningforBreedingLivestock   ̜ F   Inanimalbreeding,therapidspreadofcertaintraitswithinstocksofdomesticanimalsisofobviouscommercialimportanceandhasverylonghistoricalstanding.Artificialinseminationandembryotransfercanincreasetheeffectivereproductiveoutputofindividualelitemaleandfemaleanimalsandarewidelyusedinthelivestockindustry.Nucleartransfercloning,especiallyfromsomaticcellnuclei,couldprovideanadditionalmeansofexpandingthenumberofchosenlivestock.Theabilitytomakeidenticalcopiesofadultprizecows,sheep,andpigsisafeatureuniquetonucleartransfertechnologies,andmaywellbeusedinlivestockproduction,iftheefficienciesofadultnucleartransfercanbeimproved.Theneteffectofmultiplyingchosenanimalsbycloningwillbetoreducetheoverallgeneticdiversityinagivenlivestockline,likelywithsevereadverselong-termconsequences.Ifthistechniquebecamewidespread,effortswouldhavetobemadetoensureapoolofgeneticallydiverseanimalsforfuturelivestockmaintenance.̜  )$( ImprovedGenerationandPropagationofTransgenicLivestock   ̜ F   Thereisconsiderableinterestinbeingabletogeneticallyalterfarmanimalsbyintroductionandexpressionofgenesfromotherspecies,suchashumans.So-called"transgenicanimals"werefirstdevelopedusingmice,bymicroinjectionofDNAintothenucleusoftheegg.Thisabilitytoaddgenestoanorganismhasbeenamajorresearchtoolforunderstandinggeneregulationandforusingthemouseasamodelinstudiesofcertainhumandiseases.Ithasalsobeenappliedtootherspeciesincludinglivestock.Proposedapplicationsofthistechnologytolivestockimprovementincludethepossibleintroductionofgrowth-enhancinggenes,genesthataffectmilkqualityorwoolfibers,ordisease-resistancegenes(WardandNancarrow,1995).Butprogresshasbeenslow.Initialresultsofthemanipulationofmeatproductionbyexpressionofexcessgrowthhormoneinpigsledtoundesirablesideeffects(Pursel,etal.,1989). F   Currently,themajoractivityinlivestocktransgenesisisfocusedonpharmaceuticalandmedicalapplications.ThemilkoflivestockanimalscanbemodifiedtocontainlargeamountsofpharmaceuticallyimportantproteinssuchasinsulinorfactorVIIIfortreatmentofhumandiseasebyexpressinghumangenesinthemammarygland(Houdebine,1994).Insheepgreaterthan50percentoftheproteinsinmilkcanbetheproductofahumangene(Colman,1996).Themilkofeventransgenicmicecanyieldlarge(milligram)quantitiesofrecombinantproteins.Sincemanysuchproteinsareactiveatverylowconcentrations,itisestimatedthatproductionofhumandrugsfromtransgenicanimalscouldbeconsiderablymorecost-effectivethancurrentmethods. F   Anothermajorareaofinterestistheuseoftransgenicanimalsfororgantransplantationintohumans.Pigorgans,inmanycases,aresimilarenoughtohumanstobepotentiallyusefulinorgantransplants,ifproblemsofrejectioncouldbeovercome.Rejectioncanalreadybepartlyovercomebytheexpressionofhumancomplement(acomponentoftheimmunesystem)regulatoryproteinsintransgenicpigs.Furthertransgenicmanipulationsuchastheexpressionofhumanantigensinpigscouldalleviateorganshortagesbyminimizingoreliminatingtherejectionofpigorganstransplantedintohumans,althoughotherbarriers,suchasthepossibletransmissionofvirusesfrompigstohumans,mustbeovercome. F   Howeverthecurrentmethodofdirectlyinjectinggenesintofertilizedeggsisinefficient.Notallinjectedeggswilldevelopintotransgenicanimals,andthennotalltransgenicanimalswillexpresstheaddedgeneinthedesiredmanner.Theproductionoftransgeniclivestockisslowandexpensive.Nucleartransferwouldspeeduptheexpansionofasuccessfultransgenicline,but,perhapsmoreimportantly,itwouldallowmoreefficientgenerationoftransgenicanimalsinthefirstplace.ForeignDNA,suchasahumangene,couldbeintroducedintocelllinesincultureandcellsexpressingthetransgenecouldbecharacterizedandusedasasourceofdonornucleiforcloning,andalloffspringwouldlikelyexpressthehumangene.This,infact,wasthemotivationbehindtheexperimentsthatledtotheproductionofDolly.Ifahumangenesuchasthatforinsulincouldbeexpressedinthemammarygland,themilkofthesheepwouldbeanexcellentsourceofinsulintotreatdiabetes.  +&* GeneratingTargetedGeneAlterations    F   Themostpowerfultechnologyforgenereplacementinmammalswasdevelopedinmice.ThistechniqueaddsmanipulatedorforeignDNAtocellsinculturetoreplacetheDNApresentinthegenomeofthecells.Thusmutationsorotheralterationsthatwouldbeusefulinmedicalresearchcanbeintroducedintoananimalinadirectedandcontrolledmannerandtheireffectsstudied,aprocesscalledgenetargeting(Capecchi,1989).Thistechnologywouldbeoflimited t  use,however,withoutsomemeansoftakingthechangesgeneratedinculturedcellsandreintroducingthemintoanimals.Inmice,thiscanbeachievedbytheuseofembryonicstem(ES)cellsthatarecapableofbeingculturedindefinitelyintheundifferentiatedstate.EScellsretainthepotentialtoformallcellsoftheanimal,includingthegermcells,whenreturnedtotheenvironmentoftheearlyembryo(figure5).Asthetechniqueiscurrentlyusedinmice,thefirstgenerationofanimalsgeneratedfromtheEScellsare"chimeric,"thatistheyaremadeupofamixtureofcellsfromtwodifferentanimals.Thesemicemustthenbebredonemoretimetotransmitalteredgenestothenextgeneration.Usingthistechnique,anygeneticalterationmadeintheembryonicstemcellsinculturecanbeintroducedbackintomice(Robertson,1986). ==^#^# ====m3D9)%`|0G `(E  =y m ==== ==== ==== ==== ==== ==== ==== ==== ==== ==== ==== ==== ====̜ ==== ==== ^#^#== F Thisuseofgenereplacementandembryonicstemcelltechnologyhasbeenresponsiblefortheexplosioninthegenerationof knock-outmice,inwhichspecificgeneshavebeendeletedfromthegenome.Thesemicehavebeeninvaluableincurrentstudiestounderstandnormalgenefunctionandtoallowthegenerationofaccuratemodelsofhumangeneticdisease.Genetargetingapproachescanalsobeusedtoensurecorrecttissue-specificexpressionofforeigngenesandtosuppresstheexpressionofgenesininappropriatetissues.Ifappliedtodomesticanimals,thistechnologycouldincreasetheefficiencyoftheexpressionofforeigngenesbytargetingtheintroducedgenestoappropriateregionsofthechromosome.Itcouldalsobeusedtodirectlyalterthenormalgenesoftheorganism,whichcouldinfluenceanimalhealthandproductivityortohelp +&* developtransgenicorgansthatarelesslikelytoberejectedupontransplantation.However,todate,therearenofullyvalidatedembryonicstemcelllinesindomesticanimals.Nucleartransferfromsomaticcelllinesintoanegg,asreportedbyWilmutandcolleagues,providesapossiblealternativetotheembryonicstemcellrouteforintroductionoftargetedgenealterationsintothegermlineofanimals. F   Apartfromthefactthatembryonicstemcelllineshavenotyetbeenproducedfromfarmanimals,theotherargumentforusingnucleartransfertointroducegermlinegeneticalterationsinfarmanimalsisthatiteliminatesonegenerationofbreedingfromtheinitialchimericanimals.Thisisanimportanttimeandcostsavingfactorinfarmanimalswithlonggenerationtimesandsmalllittersize.Howeverthisfactormightnotbeasimportantasoncethought.Inmice,itturnsout,embryonicstemcellscanalsobeusedtogenerateclonedanimalscarryinggenealterationsdirectlywithouttheinitialgenerationofchimericanimals.When'tetraploid'embryosthatarenotthemselvescapableofdevelopingnormallyareusedasthehostcells,theentiremousefetuscanbederiveddirectlyfromthenormaldiploidEScells(figure6)(Nagy,etal.,1993).Althoughthisprocedureisnotyetveryefficient,itillustratestheremarkablepropertiesofEScellsandsuggeststhatsimilarapproachescouldbeappliedinotherspeciessuchasfarmanimals.̜ m3F9)%`|020 `(E0 0 Q m ^#^#               ^#^#BasicResearchonCellDifferentiation  D$X"  F   ThebasiccellularprocessesthatallowedthebirthofDollybynucleartransferusingthenucleusfromanadultsomaticdonorcellarenotwellunderstood.Ifindeedthedonorcellwasafullydifferentiatedcellandnotarare,lessdifferentiatedstemcellthatresultedinthisclonedsheep,therewillbemanyquestionstoaskabouthowthisprocessoccurred.Howthespecializedcellfromthemammaryglandwasreprogrammedtoallowtheexpressionofacompletedevelopmentalprogramwillbeafascinatingareaofstudy.Developmentalbiologistswillwanttoknowwhichgenesarereprogrammed,whentheyareexpressed,andinwhatorder.Thismight +&* shedlightonthestillpoorlyunderstoodprocessofsequentialspecializationthatmustoccurduringdevelopmentofallorganisms. F   Molecularbiologistswillalsolikelylearnmuchfromstudyinghowreprogrammingandreactivationoccurred.Whatregulatoryproteinsinthehosteggparticipatedinthereprogramming?HowdidtheseproteinsinteractwitheachotherandtheDNAsothatinactivegenesfromthemammaryglandcellsmightbeactivatedagain?Answerstothesekindsofquestionswillcontributetoouroverallunderstandingofhowcellsgrow,divide,andbecomespecialized. F   Basicresearchintothesefundamentalprocessesmayalsoleadtothedevelopmentofnewtherapiestotreathumandisease.Itisnotpossibletopredictfromwheretheessentialnewdiscoverieswillcome.However,alreadythebirthofDollyhassparkedideasaboutpotentialbenefitsthatmightberealized.Toexplorethepossibilityofthesenewtherapies,extensivebasicresearchisneeded. F   Muchofthisbasicresearchwilllikelybedoneinthemouseasthisanimaliswidelyusedbydevelopmentalbiologists,andthusagreatdealisalreadyknownaboutitsdevelopment.However,asdescribedabove,theuseofcloninginotheranimals"suchascows,pigsandsheep"byagriculturalandbiotechnologycompanieswillalsocontributetounderstandingofthebasicprocessesinvolved.Thestudyofnucleartransplantationcloninginawidevarietyofanimalswillbeveryuseful.Althoughmanyofthebasiccellularmechanismsunderlyinganimaldevelopmentarethesameinallmammals,therearesubtledevelopmentalvariationsthatoftenleadtomajortechnicaldifferencesinworkingwithaparticularspecies.Becauseatechniqueisoftenperfectedinonespeciesbeforebeingappliedtoanother,knowingwhichpartsofthetechniquesarewidelyapplicableandwhichmightneedtobeperfectedforthegivenspecieswillbeofgreatvalue.Thisbodyofresearchintoanimalsystemswillanswermanyquestionsaboutthefeasibilityofvariousnewtherapeuticapplicationsbeingproposedforhumancells.Newinnovationsintreatinghumandiseasecanbetestedinanimalsystemstodetermineifthebasicfoundationoftheideaissoundbeforeexperimentsusinghumancellswouldberequired.Thusthepathtotestingthepotentialtherapiestotreathumandisease,describedbelow,shouldinitiallygothroughtestinginanimalmodelsbeforeprogressingtohumancellresearch.  #XXXX&h#Xf9XXXPotentialTherapeuticApplicationsofNuclearTransferCloning#XXXXf9&#X&XXX  X#l!    F   Thedemonstrationthat,inmammalsasinfrogs,thenucleusofasomaticcellcanbereprogrammedbytheeggenvironmentprovidesfurtherimpetustostudiesonhowtoreactivateembryonicprogramsofdevelopmentinadultcells.Thesestudieshaveexcitingprospectsforregenerationandrepairofdiseasedordamagedhumantissuesandorgans,andmayprovidecluesastohowtoreprogramadultdifferentiatedcellsdirectlywithouttheneedforoocytefusion.Inaddition,theuseofnucleartransferhaspotentialapplicationinthefieldofassistedreproduction.̜  +&* PotentialApplicationsinOrganandTissueTransplantation    F   Manyhumandiseases,whentheyaresevereenough,aretreatedeffectivelybyorganortissuetransplantation,includingsomeleukemias,liverfailure,heartandkidneydisease.Insomeinstancestheorganrequiredisnon-vital,thatis,itcanbetakenfromthedonorwithoutgreatrisk(e.g.,bonemarrow,blood,kidney).Inothercases,theorganisobviouslyvitalandrequiredforthesurvivaloftheindividual,suchastheheart.Alltransplantationisimperfect,withtheexceptionofthatwhichoccursbetweenidenticaltwins,becausetransplantationoforgansbetweenindividualsrequiresgeneticcompatibility. F   Inprinciple,theapplicationofnucleartransfercloningtohumanscouldprovideapotentialsourceoforgansortissuesofapredeterminedgeneticbackground.Thenotionofusinghumancloningtoproduceindividualsforusesolelyasorgandonorsisrepugnant,almostunimaginable,andmorallyunacceptable.Amorallymoreacceptableandpotentiallyfeasibleapproachistodirectdifferentiationalongaspecificpathtoproducespecifictissues(e.g.,muscleornerve)fortherapeutictransplantationratherthantoproduceanentireindividual . Givencurrent   uncertaintiesaboutthefeasibilityofthis,however,muchresearchwouldbeneededinanimalsystemsbeforeitwouldbescientificallysound,andthereforepotentiallymorallyacceptable,togoforwardwiththisapproach. PotentialApplicationsinCell-basedTherapies  L`  F   Anotherpossibilityraisedbycloningistransplantationofcellsortissuesnotfromanindividualdonorbutfromanearlyembryoorembryonicstemcells;theprimitive,undifferentiatedcellsfromtheembryothatarestilltotipotent.Thispotentialapplicationwouldnotrequirethegenerationandbirthofaclonedindividual.Embryonicstemcellsprovideaninterestingmodelforsuchstudies,sincetheyrepresenttheprecursorsofallcelllineagesinthebody.Mouseembryonicstemcellscanbestimulatedtodifferentiateinvitrointoprecursorsof  theblood,neuronalandmusclecelllineages,amongothers(WeissandOrkin,1995),andtheythusprovideapotentialsourceofstemcellsforregenerationofalltissuesofthebody. F   Itmightbepossibletotakeacellfromanearlyblastomereandtreatitinsuchamannerastodirectitsdifferentiationalongaspecificpath.Bythisprocedureitmightbepossibletogenerateinthelaboratorysufficientnumbersofspecializedcells,forexamplebonemarrowstemcells,livercells,orpancreaticbeta-cells(whichproduceinsulin)fortransplantation.Ifevenasingletissuetypecouldbegeneratedfromearlyembryoniccellsbythesemethodsandusedclinically,itwouldconstituteamajoradvanceintransplantationmedicinebyprovidingcellsthataregeneticallyidenticaltotherecipient. F   Onecouldimaginetheprospectofnucleartransferfromasomaticcelltogenerateanearlyembryoandfromitanembryonicstemcelllineforeachindividualhuman,whichwouldbeideallytissue-matchedforlatertransplantpurposes.Thismightbearatherexpensiveand +&* far-fetchedscenario.Analternativescenariowouldinvolvethegenerationofafew,widelyusedandwellcharacterizedhumanembryonicstemcelllines,geneticallyalteredtopreventgraftrejectioninallpossiblerecipients. F   Theprecedingscenariosdependonusingcellsofearlyhumanembryos,generatedeitherbyinvitrofertilizationornucleartransferintoanegg.Becauseofethicalandmoralconcerns   raisedbytheuseofembryosforresearchpurposesitwouldbefarmoredesirabletoexplorethedirectuseofhumancellsofadultorigintoproducespecializedcellsortissuesfortransplantationintopatients.Itmaynotbenecessarytoreprogramterminallydifferentiatedcellsbutrathertostimulateproliferationanddifferentiationofthequiescentstemcellswhichareknowntoexistinmanyadulttissues,includingeventhenervoussystem(Gage,etal.,1995).Experimentsinthisareaarelikelytofocusmoreontheconditionsrequiredfordirectstimulationofthestemcellsinspecifictissues,thanactualuseofnucleartransfertoactivatenoveldevelopmentalprograms.Theseapproachestocellularrepairusingadultstemcellswillbegreatlyaidedbyanunderstandingofhowstemcellsareestablishedduringembryogenesis. F   Anotherstrategyforcell-basedtherapieswouldbetoidentifymethodsbywhichsomaticcellscouldbe dedifferentiatedandthen re-differentiatedalongaparticularpath.Thiswouldeliminatetheneedtousecellsobtainedfromembryos.Suchanapproachwouldpermitthegrowthofspecializedcells compatiblewithaspecificindividualpersonfortransplantation .  p Althoughatthecurrenttimethisstrategyishighlyspeculative,ongoingresearchinanimalsystemsmayidentifynewapproachesornewmoleculartargetsthatmightmakethisapproachfeasible. F   Itwillbeofgreatimportancetounderstandthroughexperimentsinanimalshowtheenvironmentoftheeggreprogramsasomaticcellnucleus.Whatcellularmechanismscanbeelucidated?Whatcomponentsareinvolvedintheseprocesses?Canwedirectcellsalongparticulardevelopmentalpathwaysinthelaboratoryandusethesecellsfortherapy?Thecapacitytogrowhumancellsofdifferentlineagesinculturewouldalsodramaticallyimproveprospectsforeffectivesomaticgenetherapy. AssistedReproduction  !  F   Anotherareaofmedicinewheretheknowledgegainedfromanimalworkhaspotentialapplicationisintheareaofassistedreproduction.Assistedreproductiontechnologiesarealreadywidelyusedandencompassavarietyofparentalandbiologicalsituations,thatis,donorandrecipientrelationships.Inmostcases,aninfertilecoupleseeksremedythrougheitherartificialinseminationorinvitrofertilizationusingspermfromeitherthemaleorananonymousdonor,aneggfromthewomanoradonor,andinsomecasessurrogacy.Inthoseinstanceswherebothindividualsofacoupleareinfertileortheprospectivefatherhasnon-functionalsperm,onemightenvisionusingcloningofonememberofthecouplesnucleitoproduceachild.  *%)  F   Althoughthisconstitutesanextensionofcurrentclinicalpractice,asidefromtheserious, moral,andethicalissuessurroundingthisapproach,therearesignificanttechnicalandmedicalcausesforcaution,someofwhichweredescribedintheresearchquestionsenumeratedabove. F   Inmostsituationsofassistedreproduction,otherthantheintentionalunionofthegametesbyinvitrotechniques,thefertilizedeggandinitialcellsoftheearlyembryoarenototherwise   manipulated.Insomerarecases,suchaspreimplantationgeneticdiagnosis,theembryoismanipulatedbytheremovalofoneoftheidenticalcellsoftheblastomeretotestitsgeneticstatus.Incontrast,ifnucleartransferweretobeusedasareproductiveoption,itwouldentailsubstantiallymoreinvasivemanipulation.Thusfar,theanimalcloningofDollyisasingularsuccess,oneseeminglynormalanimalproducedfrom277nucleartransfers.Untiltheexperimentisreplicatedtheefficiency,andeventhevalidity,oftheprocedurecannotbefullydetermined.Itislikelythatthemereactofmanipulatinganucleusandtransferringitintoaneggcoulddecreasethepercentageofeggsthatgoontodevelopandimplantnormally,aswellasincreasetherateofbirthdefects. #XXXX&#Xf9XXX@ CloningandGeneticDeterminism #XXXXf96#X&XXXԈ   F   TheannouncementofDollysparkedwidespreadspeculationaboutahumanchildbeingcreatedusingsomaticcellnucleartransfer.Muchoftheperceivedfearthatgreetedthisannouncementcenteredaroundthemisperceptionthatachildormanychildrencouldbeproducedwhowouldbeidenticaltoanalreadyexistingperson. F   Thisfearreflectsanerroneousbeliefthatapersonsgenesbearasimplerelationshiptothephysicalandpsychologicaltraitsthatcomposethatindividual.Thisbelief,thatgenesalonedetermineallaspectsofanindividual,iscalled"geneticdeterminism."Althoughgenesplayanessentialroleintheformationofphysicalandbehavioralcharacteristics,eachindividualis,infact,theresultofacomplexinteractionbetweenhisorhergenesandtheenvironmentwithinwhichtheydevelop,beginningatthetimeoffertilizationandcontinuingthroughoutlife.Associalandbiologicalbeingswearecreaturesofourbiological,physical,social,political,historical,andpsychologicalenvironments.Indeed,thegreatlessonofmodernmoleculargeneticsistheprofoundcomplexityofbothgenegeneinteractionsandgeneenvironmentinteractionsinthedeterminationofwhetheraspecifictraitorcharacteristicisexpressed.Inotherwords,therewillneverbeanotheryou. F   Whiletheconceptofcompletegeneticdeterminismiswrongandoverlysimplistic,genesdoplayamajorroleindeterminingbiologicalcharacteristicsincludingapredispositiontocertainϜdiseases.Moreover,theexistenceoffamiliesinwhichmanymembersareaffectedbythesediseasessuggestthatthereisasinglegenethatispasseddownwitheachgenerationthatcausesthedisease.Whensuchadiseasegeneisidentified,scientistsoftensaytheyhave"clonedthegenefor"breastcancer,forinstance,implyingadirectcauseandeffectofgeneanddisease. Indeed,therecenteffortsoftheHumanGenomeProjecthaveledtotheisolationofalarge *%) numberofgenesthataremutatedinspecificdiseases,suchasDuchenneMuscularDystrophy,andcertaintypesofbreastandcoloncancer.  F   However,recentscientificfindingshaverevealedthata onegene,onediseaseapproachisfartoosimplistic.Evenintherelativelysmalllistofgenescurrentlyassociatedwithaspecificdisease,knowingthecompleteDNAsequenceofthegenedoesnotallowascientisttopredictifagivenpersonwillgetthedisease.Forexample,inbreastcancertherecanbemanydifferentchangesintheDNA,andforsomespecificmutationsthereisacalculatedriskofdevelopingthedisease,whileforotherchangestheriskisunknown.Evenwhenaspecificgeneticchangeisidentifiedthat"causes"thediseaseinsomepeople,othersmaybefoundwhohavethesamechangebutdonotgetthedisease.Thisisbecauseotherfactors,eithergeneticorenvironmental,arealteredthatmaskorcompensatefor"the"diseasegene.Thusevenwiththemostsophisticatedunderstandingofgenes,onecannotdeterminewithcertaintywhatwillhappentoagivenpersonwithasinglechangeinasinglegene. F   Onceagain,thereasonrigidgeneticdeterminismisfalseisthatgenesinteractwitheachotherandwiththeenvironmentinextremelycomplexways.Forexample,thelikelihoodofdevelopingcoloncancer,adiseasewithastronghereditarycomponentandforwhichresearchershaveidentifiedasingle"causative"gene,isalsostronglyinfluencedbydiet.Whenoneconsidersahumantraitthatisdeterminedbymultiplegenes,thesituationbecomesevenmorecomplex.Thenumberofinteractionsbetweengenesandenvironmentincreasesdramatically.Infact,theabilitytopredictwhatapersonwillbelikeknowingonlytheirgenesbecomesvirtuallyimpossiblebecauseitisnotpossibletoknowhowtheenvironmentandchancefactorswillinfluencetheoutcome. F   ThustheideathatonecouldmakethroughsomaticcellnucleartransferateamofMichaelJordans,aphysicsdepartmentofAlbertEinsteins,oranoperachorusofPavarottis,issimplyfalse.Knowingthecompletegeneticmakeupofanindividualwouldnottellyouwhatkindofpersonthatindividualwouldbecome.Evenidenticaltwinswhogrowuptogetherandthussharethesamegenesandasimilarhomeenvironmenthavedifferentlikesanddislikes,andcanhaveverydifferenttalents.Theincreasinglysophisticatedstudiescomingoutofhumangeneticsresearchareshowingthatthebetterweunderstandgenefunction,thelesslikelyitiswewilleverbeabletoproduceatwillapersonwithanygivencomplextrait.  #XXXX&#Xf9XXXConclusions #XXXXf9#X&XXX D$X"    F   Theterm clonehasmanymeaningsbutinitssimplestandmostscientificsenseitmeansthemakingofidenticalcopiesofmolecules,cells,tissues,andevenentireanimals.ThelatestnewsaboutcloningDollythesheepinvolvedsomaticcellnucleartransplantcloning.Inthisprocessthenucleusfromanadultsomaticcellistransplantedintoanenucleatedovumtoproduceadevelopinganimalthatisa delayedgenetictwinoftheadult.  *%)  F   Therearemanyapplicationsthatnucleartransfercloningmighthaveforbiotechnology, livestockproduction,andnewmedicalapproaches.Workwithembryonicstemcellsandgeneticmanipulationofearlyembryosinanimalspecies(includingnucleartransfer)isalreadyprovidingunparalleledinsightsintofundamentalbiologicalprocessesandpromisestoprovidegreatpracticalbenefitintermsofimprovedlivestock,improvedmeansofproducingpharmaceuticalproteins,andprospectsforregenerationandrepairofhumantissues. F   However,thepossibilityofusinghumancloningforthepurposesofcreatinganewindividualentailssignificantscientificuncertaintyandmedicalriskatthistime.Potentialrisksincludethoseknowntobeassociatedwiththemanipulationofnucleiandeggsandthoseyetunknown,suchastheeffectsofaging,somaticmutation,andimproperimprinting.Theseeffectscouldresultinhighratesoffailedattemptsatpregnancyaswellastheincreasedlikelihoodofdevelopmentallyandgeneticallyabnormalembryos.      #XXXX&G#Xf9XXXReferences#XXXXf90##X&XXX   "0 F  Baron,M.H.,andT.Maniatis,"Rapidreprogrammingofglobingeneexpressionintransient F^#F^#  F   heterokaryons,"Cell46:591-602,1986.  0 F  Blau,H.M.,"Differentiationrequirescontinuousactivecontrol,"AnnualReviewofBiochemistry F^#F^#  F   61:1213-1230,1992.0 F  Blau,H.M.,Pavlath,G.K.,Hardeman,E.C.,Chiu,C.P.,Silberstein,L.,Webster,S.G.,Miller, F^#F^#  F   S.C.,andC.Webster,"Plasticityofthedifferentiatedstate,"Science230:758-766,1985. 8 L  0 F  Braude,P.,Bolton,V.,andS.Moore,"Humangeneexpressionfirstoccursbetweenthefour-and @P#  F^#F^#  F   eight-cellstagesofpreimplantationdevelopment,"Nature332:459-461,1988.   0 F  Briggs,R.,andT.J.King,"Transplantationoflivingnucleifromblastulacellsintoenucleated F^#F^#  F   frog'seggs,"ProceedingsoftheNationalAcademyofSciences(USA)38:455-463,1952.   0 F  Campbell,K.H.,Loi,P.,Cappai,P.,andI.Wilmut,"ImproveddevelopmenttoblastocystofovineϜ  nucleartransferembryosreconstructedduringthepresumptiveS-phaseofenucleated F^#F^#  F   activatedoocytes,"BiologyofReproduction50:1385-1393,1994. p 0 F  Campbell,K.H.,McWhir,J.,Ritchie,W.A.,andI.Wilmut,"Sheepclonedbynucleartransfer F^#F^#  F   fromaculturedcellline,"Nature380:64-66,1996. 4H 0 F  Capecchi,M.R.,"Thenewmousegenetics:alteringthegenomebygenetargeting,"Trendsin  F^#F^#  F   Genetics5:70-76,1989.   0 F  Cheong,H.T.,Takahashi,Y.,andH.Kanagawa,"Birthofmiceaftertransplantationofearly F^#F^#  F   cell-cycle-stageembryonicnucleiintoenucleatedoocytes,"BiologyofReproduction   F   48:958-963,1993.  0 F  Chiu,C.P.,andC.B.Harley,"Replicativesenescenceandcellimmortality:Theroleoftelomeres F^#F^#  F   andtelomerase,"ProceedingsoftheSocietyforExperimentalBiologyandMedicine l"   F   214:99-106,1997.0 F  Collas,P.,andF.L.Barnes,"Nucleartransplantationbymicroinjectionofinnercellmassand F^#F^#  F   granulosacellnuclei,"MolecularReproductionandDevelopment38:264-267,1994. &0!$ 0 F  Collas,P.,Balise,J.J.,andJ.M.Robl,"Influenceofcellcyclestageofthedonornucleuson F^#F^#  F   developmentofnucleartransplantrabbitembryos,"BiologyofReproduction46:492-500, (#'  F   1992.̜  *%) Colman,A.,"Productionofproteinsinthemilkoftransgeniclivestock:Problems,solutions,and F   successes,"AmericanJournalofClinicalNutrition63:639S-645S,1996.  0 F  Fundele,R.H.,andM.A.Surani,"Experimentalembryologicalanalysisofgeneticimprintingin F^#F^#  F   mousedevelopment,"DevelopmentalGenetics15:515-522,1994.  0 F  Fulka,J.,First,N.L.,andR.M.Moor,"Nucleartransplantationinmammals:remodelingof F^#F^#  F   transplantednucleiundertheinfluenceofmaturationpromotingfactor,"BioEssays (#  ` t  F   18:835-840,1996.0 F  Graham,C.F."Thefusionofcellswithone-andtwo-cellmouseembryos,"WistarInstitute$8 F^#F^#  F   SymposiumMonographs9:19-35,1969. $  0 F  Gage,F.H.,Ray,J.,andL.J.Fisher,"Isolation,characterization,anduseofstemcellsfromthe (#  F^#F^#  F   CNS,"AnnualReviewofNeuroscience18:159-192,1995.   0 F  Gurdon,J.B."Thedevelopmentalcapacityofnucleitakenfromintestinalepitheliumcellsof F^#F^#  F   feedingtadpoles,"JournalofEmbryologyandExperimentalMorphology10,622-640,   F   1962.0 F  Gurdon,J.B.,Laskey,R.A.,andO.R.Reeves,"Thedevelopmentalcapacityofnuclei F^#F^#  F   transplantedfromkeratinizedskincellsofadultfrogs,"JournalofEmbryologyand H\  F   ExperimentalMorphology34:93-112,1975. 4H 0 F  Houdebine,L.M.,"Productionofpharmaceuticalproteinsfromtransgenicanimals,"Journalof  F^#F^#  F   Biotechnology34:269-287,1994.   0 F  Latham,K.E.,Garrels,J.I.,andD.Solter,"Alterationsinproteinsynthesisfollowing F^#F^#  F   transplantationofmouse8-cellstagenucleitoenucleated1-cellembryos,"   F   DevelopmentalBiology163:341-350,1994.  0 F  Mantell,L.L.,andC.W.Greider,"Telomeraseactivityingermlineandembryoniccellsof F^#F^#  F   Xenopus,"EMBOJournal13:3211-3217,1994. l"  0 F  McGrath,J.,andD.Solter,"Inabilityofmouseblastomerenucleitransferredtoenucleated F^#F^#  F   zygotestosupportdevelopmentinvitro,"Science226:1317-1319,1984. 0%D # 0 F  Nagy,A.,Rossant,J.,Nagy,R.,Abramow-Newerley,W.,andJ.C.Roder,"Derivationof F^#F^#  F   completelycellculture-derivedmicefromearlypassageembryonicstemcells," '#&  F   ProceedingsoftheNationalAcademyofSciences(USA)90:8424-8428,1993. (#' 0 F  Prather,R.S.,Sims,M.M.,andN.L.First,"Nucleartransplantationinearlypigembryos,"Biology*%)F^#F^#  F   ofReproduction41:414-418,1989. +&* Ї0 F  Prather,R.S.,andN.L.First,"Cloningembryosbynucleartransfer,"JournalofReproductionF^#F^#  F   andFertility.Supplement41:125-134,1990.  0 F  Pursel,V.G.,Pinkert,C.A.,Miller,K.F.,Bolt,D.J.,Campbell,R.G.,Palmiter,R.D.,Brinster, F^#F^#  F   R.L.,andR.E.Hammer,"Geneticengineeringoflivestock,"Science244:1281-1288,   F   1989.0 F  Robertson,E.J.,"Pluripotentialstemcelllinesasarouteintothemousegermline,"Trendsin` tF^#F^#  F   Genetics2:9-13,1986. L ` 0 F  RossantJ.,andR.A.Pederson,ExperimentalApproachestoMammalianEmbryonicDevelopment$8 F^#F^#  F   (Cambridge:CambridgeUniversityPress,1986). $  0 F  Schultz,R.M.,"Regulationofzygoticgeneactivationinthemouse,"BioEssays15:531-538, F^#F^#  F   1993.0 F  Sims,M.,andN.L.First,"Productionofcalvesbytransferofnucleifromculturedinnercellmass F^#F^#  F   cells,"ProceedingsoftheNationalAcademyofSciences(USA)91:6143-6147,1994.  0 F  Smith,L.C.,andI.Wilmut,"Influenceofnuclearandcytoplasmicactivityonthedevelopmentin F^#F^#  F   vivoofsheepembryosafternucleartransplantation,"BiologyofReproduction \p  F   40:1027-1035,1989.0 F  Solter,D.,"Differentialimprintingandexpressionofmaternalandpaternalgenomes,"Annual 4F^#F^#  F   ReviewofGenetics22:127-146,1988.    0 F  Stice,S.L.,andC.L.Keefer,"Multiplegenerationalbovineembryocloning,"BiologyofF^#F^#  F   Reproduction48:715-719,1993.  0 F  Stice,S.L.,Keefer,C.L.,andL.Mathews,"Bovinenucleartransferembryos:Oocyteactivation F^#F^#  F   priortoblastomerefusion,"MolecularReproductionandDevelopment38:61-68,1994.   0 F  Ward,K.A.,andC.D.Nancarrow,"Thecommercialandagriculturalapplicationsofanimal F^#F^#  F   transgenesis,"MolecularBiotechnology4:167-178,1995. X#l! 0 F  Weintraub,H.,"TheMyoDfamilyandmyogenesis:redundancy,networks,andthresholds,"Cell0%D #F^#F^#  F   75:1241-1244,1993.0 F  Weiss,M.J.,andS.H.Orkin,"Invitrodifferentiationofmurineembryonicstemcells:New F^#F^#  F   approachestooldproblems,"JournalofClinicalInvestigation97:591-595,1995. (#' Willadsen,S.M.,"Nucleartransplantationinsheepembryos,"Nature320:63-65,1986. *%)  +&* 0 F  Wilmut,I.,Schnieke,A.E.,McWhir,J.,Kind,A.J.,andK.H.Campbell,"Viableoffspring F^#F^#  F   derivedfromfetalandadultmammaliancells,"Nature385:810-813,1997.  0 F  YangX.,Jiang,S.,Kovacs,A.,andR.H.Foote,"Nucleartotipotencyofculturedrabbitmorulae @#  F^#F^#  F   tosupportfull-termdevelopmentfollowingnucleartransfer,"BiologyofReproduction   F   47:636-643,1992.#XXXX&|##    4,H'* U9ћ7XXdXXd7Xf9XXXV^XXf9  Ӝ ChapterThree  RELIGIOUSPERSPECTIVES   A- ) xdE0A  )P# ^ V^O#  ^#X&X P#Overthelasttwenty-fiveyearsreligiousthinkershavediscussedtheprospectofhumancloningin   thecontextoflong-standingreligioustraditionsthatofteninfluenceandguidecitizens'responsestonewtechnologies.Religiouspositionsonhumancloningarepluralisticintheirpremises,modesofargument,andconclusions.Nevertheless,severalmajorthemesareprominentinJewish,RomanCatholic,Protestant,andIslamicpositions,includingresponsiblehumandominionovernature,humandignityanddestiny,procreation,andfamilylife.Somereligiousthinkersarguethatcloningahumantocreateachildwouldbeintrinsicallyimmoralandthuscouldneverbemorallyjustified;theyusuallyproposeabanonsuchhumancloning.Someotherreligiousthinkerscontendthathumancloningtocreateachildcouldbemorallyjustifiedundersomecircumstancesbutholdthatitshouldbestrictlyregulatedinordertopreventabuses.#XXXX&Q#Xf9XXX Oc  Л#XXXXf9GU#X&XXXԜ@>>********** !5 МMediareportsoftendepictthedebateovertheprospectsofcloninghumansasaclassicalconfrontationbetweenscienceandreligion.Thisdepictionismisleading.Notallargumentsagainstcloninghumansarereligious,andnotallreligiousthinkersopposethecloningofhumansinallcircumstances.Furthermore,manycontendthatthepossibilityofcloninghumansoffersanopportunityforsubstantivedialoguebetweenscientistsandtheologians.Probingtheintersectionsofethics,science,andtheologycanoffermutualenrichment.Scientistscanseehowresearchingeneticsandbiologyraisestheologicalquestions,whiletheologianscanconsiderwhetherandhowreligiousconvictionscanaccommodatenewscientificknowledge(Gustafson,1994).Therefore,theCommissionsoughttodeterminethepositionsonhumancloningviasomaticcellnucleartransferheldbyavarietyofreligiousthinkersandtheargumentstheyoffertosupporttheirpositions.TheCommissionwasinterestedinreligiousargumentsandconclusionsabouthumancloningbecausereligioustraditionsinfluenceandguidemanycitizensresponsestovariousissuesinbiomedicine,includingsuchnoveldevelopmentsashumancloning.̜ F   Forpurposesofrecommendingpublicpolicyinademocraticsociety,theCommissionwasalsointerestedintheextenttowhichmoralargumentsinvariousreligioustraditionsrestonpremisesaccessibletoothersoutsidethosetraditions.Sometimesreligiousthinkersappealtocategoriessuchas nature, reason, basichumanvalues,and familyvaluesthatmayspeaktocitizensoutsidetheirtraditionsbecausethesecategoriesdonotnecessarilydependsolelyonparticularfaithcommitments,scripture,revelation,orreligiousauthority.Suchcategoriesmaythereforecontributetoabroadersocietaldiscussionoftheethicalargumentsforandagainstcloninghumans,whichareexaminedfurtherinthefollowingchapter.Indeed,NBACwasinterestedindeterminingwhethervariousreligioustraditionsandsecularapproachesachievea  A*U%) roughconsensusaboutappropriatepublicpolicytowardcreatingchildrenthroughsomaticcellnucleartransferatthistime. F   Finally,theviewsofawiderangeofcommunities,includingreligiouscommunities,areimportantindeterminingwhetherpoliciesunderconsiderationarefeasibleandwhethertheirsocialbenefitsoutweightheirsocialcosts.Forexample,aparticularpolicymaynotbefeasible,andmayevenbecounterproductive,ifitengendersvigorous,widespread,andsustainedmoralobjection. F   NBACsolicitedoralandwrittenpresentationsfromscholarsinseveralreligioustraditions,contractedforascholarlyanalysisoftheviewsoftheseandotherreligioustraditions,#XXXX&U#X&XXXV !=  5      #XXXX&a#X&XXX $8  andreceivedpublictestimonyandwrittensubmissionsfromvariousotherindividualsandgroupswithreligiousorientations.Whatfollowsbuildsonthesematerialsandpresentssomeofthekeythemesinseveralwesternreligiousinterpretationsandevaluationsofcloninghumans.ThischapterispresentedinthespiritofsustaininganationaldialoguebutalsoincompleteawarenessthattheCommissionmaynothavefullyunderstoodthetraditionsdescribed.(ThischapterconcentratesonJewish,RomanCatholic,Protestant,andIslamicviews;adiscussionofotherreligiousviewsappearsinthecommissionedpaperbyCourtneyCampbell.) #XXXX&b#Xf9XXX  ReligionandHumanCloning:AnHistoricalOverview#XXXXf9d#X&XXX  p e F   Itispossibletoidentifyfourrecentoverlappingperiodsinwhichtheologiansandotherreligiousthinkershaveconsideredthescientificprospectsandethicsofthecloningofhumans.Thefirstphase,whichbeganinthemid1960sandcontinuedintotheearly1970s,wasshapedbyacontextofexpandedchoicesandcontrolofreproduction(e.g.,theavailabilityofthebirthcontrolpill),theprospectsofalternative,technologicallyassistedreproduction(e.g.,invitro   fertilization[IVF]),andtheadvocacybysomebiologistsandgeneticistsofcloning preferredgenotypes,which,intheirview,wouldavoidoverloadingthehumangenepoolwithgenesthatarelinkedtodeleteriousoutcomesandthatcouldplacethesurvivalofthehumanspeciesatrisk. F   Severalprominenttheologiansengagedintheseinitialdiscussionsofhumangeneticmanipulationandcloning,includingCharlesCurran,BernardHring,RichardMcCormick,andKarlRahnerwithinRomanCatholicism,andJosephFletcherandPaulRamseywithinProtestantism.Thediametricallyopposedpositionsstakedoutbythelasttwotheologiansgaveanearlysignalofthewiderangeofviewsthatarestillexpressedbyreligiousthinkers. F   JosephFletcheradvocatedexpansionofhumanfreedomandcontroloverhumanreproduction.Heportrayedthecloningofhumansasoneofmanypresentandprospectivereproductiveoptionsthatcouldbeethicallyjustifiedbysocietalbenefit.Indeed,forFletcher,asa '#& methodofreproduction,cloningwaspreferabletothe geneticrouletteofsexualreproduction.Heviewedlaboratoryreproductionas radicallyhumanbecauseitisdeliberate,designed,chosen,andwilled(Fletcher,1971;1972;1974;1979). F   Bycontrast,PaulRamseyportrayedthecloningofhumansasa borderlineormoralboundarythatcouldbecrossedonlyatriskofcompromisetohumanityandtobasicconceptsofhumanprocreation.Cloningthreatenedthree horizontal(personperson)andtwo vertical(personGod)bordercrossings.First,clonalreproductionwouldrequiredirectedormanagedbreedingtoservethescientificendsofacontrolledgenepool.Second,itwouldinvolvenontherapeuticexperimentationontheunborn.Third,itwouldassaultthemeaningofparenthoodbytransforming procreationinto reproductionandbyseveringtheunitiveend(expressingandsustainingmutuallove)andtheprocreativeendofhumansexualexpression.Fourth,thecloningofhumanswouldexpressthesinofprideorhubris.Fifth,itcouldalsobeconsideredasinofselfcreationashumansaspiretobecomea manGod(Ramsey,1966;1970). F   Aseconderaoftheologicalreflectiononcloninghumansbeganin1978,ayearthatwasnotablefortwoevents,thebirthinBritainofthefirstIVFbaby,LouiseBrown,andthepublicationofDavidRorvik'sInHisImage,anaccountalleging(falsely)thecreationofthefirst  clonedhumanbeing(Rorvik,1978).ChristiantheologiansconcentratedmoreontheethicalissuesraisedbyIVF,whileJewishscholars,suchasSeymourSiegelandFredRosner,alsodirectedattentiontocloninghumans,andwereneitherassupportiveasFletchernorascriticalasRamsey.TheyinsteadindicatedtheneedformoreextensivediscussionofthistopicwithintheJewishcommunity. F   Thisperiodalsowitnessedthebeginningofformalecclesiasticalinvolvementwithquestionsofgeneticmanipulation.In1977theUnitedChurchofChristproducedastudybookleton GeneticManipulation,whichappearstobetheearliestreferencetohumancloningamongProtestantdenominationalliterature(Lynn,1977).Itprovidedageneraloverviewofthescienceandethicsofcloninghumansbutstoppedshortofaspecifictheologicalverdict. F   ProtestantorganizedecumenicalbodiessuchastheWorldCouncilofChurches(1975,1982,1989)andtheNationalCouncilofChurchesofChrist(1980,1983,1986),aswellassomeindividualdenominations,issuedresolutionsorpositionstatementsthatcautiouslyendorsedgeneticinterventionsfortherapeuticpurposes.Inaddition,in1979,concernsaboutgeneticengineeringexpressedbyJewish,Protestant,andRomanCatholicleadersledPresidentJimmyCartertoaskthePresidentsCommissionfortheStudyofEthicalProblemsinMedicineandBiomedicalandBehavioralResearchtoexaminethescientific,ethical,andsocialissuesofgenesplicing(President'sCommission,1982).ThePresidentsCommissionaddressedtheaspectsof geneticengineeringthatuserecombinantDNAtechnologytotreatdisease,butitdidnotaddressotherproceduresoftenencompassedbythephrase,suchasIVForcloningorganisms. F   Thediscussionsofthe1970scontinuedintothe1980swithparticularattentiontoIVF,artificialinseminationbydonor,andsurrogacy.Thesetechniqueschallengedtraditionalnotions +&* ofthefamilybyseparatinggeneticandrearingfatherhoodandgenetic,gestational,andrearingmotherhood,aswellasraisingquestionsaboutwhetherthecontractualandcommercialtiesinmanyofthesearrangementswereinimicaltotraditionalreligiousviewsofthefamily. F   Athirderaofreligiousdiscussionbeganin1993withthereportfromGeorgeWashingtonUniversityoftheseparationofcellsinhumanblastomerestocreatemultiple,geneticallyidenticalembryos.TheRomanCatholicChurchexpressedvigorousoppositiontotheprocedure,andaVaticaneditorialdenouncedtheresearchas intrinsicallyperverse.Catholicmoraltheologiansinvokednormsofindividuality,dignity,andwholenessincondemningthisresearch(McCormick,1993;1994;Shannon,1994).WhilemanyConservativeProtestantscholarsheldthatthisresearchcontravenedbasicnotionsofpersonhoodsuchasfreedom,thesanctityoflife,andtheimageofGod,someotherProtestantscholarsnoteditspotentialmedicalbenefitsandadvocatedcarefulregulationratherthanprohibition. F   ThefourthandmostrecentstageofreligiousdiscussionhascomeinthewakeofthesuccessfulcloningofDollythesheepthroughthesomaticcellnucleart