Cre-LoxP系统在转基因小鼠上的应用策略

Cre/LoxP(,400038)Cre/LoxP.,/DNA,,Cre,../Cre,,,,,Q55,Q52,Q7,Q95(transgenicmice,TgM)(embryonicstem,ES).TgM,,(misexpress),;,(nullallele),ES,,[1].,.,TgM,,TgM;(knockout)/,,.Cre/LoxP,.Cre/LoxP,,.1Cre/LoxPDNA.,Int(integrasefamily)resolvase2invertasefamily.CreP1cre38ku,Int.P134bpLoxP,LoxP,(,DNA):a1(1a);b1(1b);c1(1c).,Cre,;.Cre,;Cre/LoxPDNA[1,2].1CreP:;#:LoxP;a,b,c,d:.2,TgM.,Tel:[phone],E2mail:[e-mail]:1999204205,:19992082025322000;27(3)Prog.Biochem.Biophys.,,,TgM.,TgM,,,,.,TgM,(dormanttransgene),,,,.LoxP2CATSTOP,,TgM,CAT,TgM,CreTgM,CreSTOP,(2).Lakso[3]LoxP2STOPSV40T2Ag,TgM,SV40T2Ag,CreTgM,CreSTOP,SV40T2Ag,TgM.Wakita[4],(HCV)TgM,HCV.2CreSTOP:;1.3,,,knockout/,,.,knockout:,,.,(neo)(HSVtK)(positiveandnegativeselection),;,,(neo);,.Cre/LoxP(condi2tionalgenetargeting).DNALoxP,ES,CreTgM,Cre,knockout(3a).Gu[5]TgMDNAT.Cre[6][7][8].,Cre,,,.Cre/LoxP,,(3b).,3,,,ESG418,,ESCre,neo.,LoxP2neoES,CreTgM,.CreLoxP(tagged)632Prog.Biochem.Biophys.2000;27(3)CreTgM,knockout.LoxPESknockout,.,Lobe[9],Cre,.4/DNA,.Cre,/,.,Cre.(tetracycline,Tet)Tet2off/Tet2on,Tet2off(tetracyclinerepressor,TetR)(HSV)VP16C130,(tetracyclinetranscriptionalactivator,tTA);(tetracyclineresis2tanceoperon,TetO)(cytomegalovirus,CMV)(tetracyclineresponseelement,TRE),cre.tTACreTgM;LoxP,Tet,TetTetR,tTA,TetO,TREcre;Tet,tTATRE,TREcre,Cre,/.Stonge[10],TgM.,tet,(ecdysone),RU486(chemicalinducerofdimerization,CIDs),,[11].,Cre,Cre(ER)(LBD)Cre.Brocard[12].ERLBDCre(Cre2ER).Tamoxifin.,CreLBDCre2ERt,,ERt,.CMVCre2ERtTgM,Tamoxifin,Cre.Schwenk[13]BEu/Psv40Cre2ERt,,BCre2ERtTgM,Tamoxifin,Cre,Cre.,.,Cre/LoxP.,/;Cre,/.,.Cre/LoxP,:a1Cre,,.b1FLP,.c1.,Cre/LoxPTgM(http//).,,7322000;27(3)Prog.Biochem.Biophys..1SauerB.InduciblegenetargetinginmiceusingtheCre/Loxsystem.Methods,1998,14(4):3813922GuoF,GopaulDN,DuyneGD.StructureofCrerecombinasecomplexedwithDNAinasite2specificrecombinationsynapse.Nature,1997,389(6646):40463LaksoM,SauerB,MosingerB,etal.Targetedoncogeneactivationbysitespecificrecombinationintransgenicmice.ProcNatlAcadSciUSA,1992,89(14):623262364WakitaT,TayaC,KatsumeA,etal.EfficientconditionaltransgeneexpressioninhepatitisCviruscDNAtransgenicmicemediatedbytheCre/loxPsystem.JBiolChem,1998,273(15):900190065GuH,MarthJD,OrbanPC,etal.DeletionofDNApoly2merasegenesegmentinTcellusingcelltype2specificgenetargeting.Science,1994,265(5168):1031066KellendonkC,TrocheF,CasanovaE,etal.Induciblesite2specificrecombinationinthebrain.JMolBiol,1999,285(1):1751827WagnerKU,WallRJ,StOngeL,etal.Cre2mediatedgenedeletioninthemammarygland.NucleicAcidsRes,1997,25(21):432343308ShibataH,ToyamaK,ShioyaH,etal.RapidcolorectaladenomaformationinitiatedbyconditionaltargetingoftheApcgene.Science,1997,278(5335):1201239LobeCG,KoopKE,KreppnerW,etal.Z/AP,adoublereporterforcre2mediatedrecombination.DevBiol,1999,208(2):28129210StongeL,UrthPA,GrussP.TemporalcontroloftheCrerecombinaseintransgenicmicebyatetracyclineresponsivepromoter.NucleicAcidsRes,1996,24(19):3875387711SaezE,NoD,WestA,etal.Induciblegeneexpressioninmammaliancellsandtransgenicmice.CurrOpinBiotech,1997,8(3):60861612BrocardJ,WarotX,WendlingO,etal.Spatio2temporallycon2trolledsite2specificsomaticmutagenesisinthemouse.ProcNatlAcadSciUSA,1997,94(26):145591456313SchwenkF,KuhnR,AngrandPO,etal.Temporallyandspatiallyregulatedsomaticmutagenesisinmice.NucleicAcidsRes,1998,26(6):14271432StrategiesandApplicationsoftheCre/LoxPSysteminTransgenicMice.ZHUHuan2Zhang,SHIJing2Quan(DepartmentofPathology,ThirdMilitaryMedicalUniversity,Chongqing400038,China).AbstractCresite2specificDNArecombinasesystemhasbeendevelopedasanovelpowerfultoolsformanipulatingDNAbothinvitroandinvivo.Ithasbeenusedintransgenicmicetoinducesite2specificDNArecombinationleadingtogeneexpressionordeletion/mutationnotonlyinatissue2specificoratacertainstageofdevelopment,bycombiningwithinduciblesystemsforcontrollingCreexpressionorfunction,butalsointemporallyandspatiallymanner.Theserecombination2basedstrategiesarelikelytohaveaprofoundimpactonstudyofgenefunctionandthegenerationofanimalmodelsofhumandiseases.Keywordsintegrase/Cre,recombination,geneexpressionregulation,transgene,genetargeting,mice(,,200031)Aequoreavictoria(greenfluorescentprotein,GFP).(max=395nm,479nm).GFP.GFP.GFP.,,,Q785(greenfluorescentprotein,GFP).,GFP.:,[1].(Aequoreavictoria)Tel:[phone],E2mail:[e-mail]:1999204208,:1999208224832Prog.Biochem.Biophys.2000;27(3)