Generation of Knockout Rats with X-Linked Severe Combined Immunodeficiency (X-SCID) Using Zinc-Finger Nucleases 英文参考文献.docVIP

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Generation of Knockout Rats with X-Linked Severe Combined Immunodeficiency (X-SCID) Using Zinc-Finger Nucleases 英文参考文献.doc

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Generation of Knockout Rats with X-Linked Severe Combined Immunodeficiency (X-SCID) Using Zinc-Finger Nucleases 英文参考文献

GenerationofKnockoutRatswithX-LinkedSevere CombinedImmunodeficiency(X-SCID)UsingZinc-Finger Nucleases TomojiMashimo1*,AkikoTakizawa1,BirgerVoigt1,KazutoYoshimi1,HiroshiHiai2,TakashiKuramoto1, TadaoSerikawa1 1InstituteofLaboratoryAnimals,GraduateSchoolofMedicine,KyotoUniversity,Kyoto,Japan,2ShigaMedicalCenterforAdultDisease,Moriyama,Japan Abstract Background: Althoughtherat is extensively used as alaboratory model, theinability toutilizegerm line-competent rat embryonicstem(ES)cellshasbeenamajordrawbackforstudiesthataimtoelucidategenefunctions.Recently,zinc-finger nucleases (ZFNs) were successfully used to create genome-specific double-stranded breaks and thereby induce targeted genemutationsinawidevarietyoforganismsincludingplants,drosophila,zebrafish,etc. Methodology/PrincipalFindings:WereporthereonZFN-inducedgenetargetingoftheratinterleukin2receptorgamma(Il2rg) locus,whereorthologoushumanandmousemutationscauseX-linkedseverecombinedimmunedeficiency(X-SCID).Co-injection ofmRNAsencodingcustom-designedZFNsintothepronucleusoffertilizedoocytesyieldedgeneticallymodifiedoffspringatrates greaterthan20%,whichpossessedawidevarietyofdeletion/insertionmutations.ZFN-modifiedfoundersfaithfullytransmitted theirgeneticchangestothenextgenerationalongwiththeseverecombinedimmunedeficiencyphenotype. ConclusionsandSignificance:TheefficientandrapidgenerationofgeneknockoutratsshowsthatusingZFNtechnologyis anewstrategyforcreatinggene-targetedratmodelsofhumandiseases.Inaddition,theX-SCIDratsthatwereestablished in this study will be valuable in vivo tools for evaluating drug treatment or gene therapy as well as model systems for examiningthetreatmentofxenotransplantedmalignancies. Citation:MashimoT,TakizawaA,VoigtB,YoshimiK,HiaiH,etal.(2010)GenerationofKnockoutRatswithX-LinkedSevereCombinedImmunodeficiency(X- SCID)UsingZinc-FingerNucleases.PLoSONE5(1):e8870.doi:10.1371/journal.pone.0008870 Editor:EllenA.A.Nollen,UniversityMedicalCenterGroningen,Netherlands ReceivedNovember12,2009;Accepted