Silencing Inhibits Cre-Mediated Recombination of the ZAP and ZEG Reporters in Adult Cells 英文参考文献.docVIP

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Silencing Inhibits Cre-Mediated Recombination of the ZAP and ZEG Reporters in Adult Cells 英文参考文献.doc

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Silencing Inhibits Cre-Mediated Recombination of the ZAP and ZEG Reporters in Adult Cells 英文参考文献

SilencingInhibitsCre-MediatedRecombinationofthe Z/APandZ/EGReportersinAdultCells MichaelA.Long,FabioM.V.Rossi* TheBiomedicalResearchCentre,UniversityofBritishColumbia,Vancouver,BritishColumbia,Canada Abstract Background: The Cre-loxPsystem has been used toenabletissue specific activation,inactivation andmutationof many genesinvivoandhastherebygreatlyfacilitatedthegeneticdissectionofseveralcellularanddevelopmentalprocesses.In suchstudies,Cre-reporterstrains,whichcarryaCre-activatedmarkergene,arefrequentlyutilizedtovalidatetheexpression profileofCretransgenes,toactasasurrogatemarkerforexcisionofasecondallele,andtoirreversiblylabelcellsforlineage tracingexperiments. Principal Findings: We have studied three commonly used Cre-reporter strains, Z/AP, Z/EG and R26R-EYFP and have demonstratedthatalthougheachreportercanbereliablyactivatedbyCreduringearlydevelopment,exposuretoCrein adulthematopoieticcellsresultsinamuchlowerfrequencyofmarker-positivecellsintheZ/APorZ/EGstrainsthaninthe R26R-EYFPstrain.InmarkernegativecellsderivedfromtheZ/APandZ/EGstrains,thetransgenicpromoterismethylated andCre-mediatedrecombinationofthelocusisinhibited. Conclusions:TheseresultsshowthattheefficiencyofCre-mediatedrecombinationisnotonlydependentonthegenomic contextofagivenloxP-flankedsequence,butalsoonstochasticepigeneticmechanismsunderlyingtransgenevariegation. Furthermore,ourdatahighlightsthepotentialshortcomingsofutilizingtheZ/APandZ/EGreportersassurrogatemarkers ofexcisionorinlineagetracingexperiments. Citation: Long MA, Rossi FMV (2009) Silencing Inhibits Cre-Mediated Recombination of the Z/AP and Z/EG Reporters in Adult Cells. PLoS ONE 4(5): e5435. doi:10.1371/journal.pone.0005435 Editor:SebastianD.Fugmann,NationalInstituteonAging,UnitedStatesofAmerica ReceivedFebruary5,2009;AcceptedApril12,2009;PublishedMay5,2009 Copyright: ? 2009 Long, Rossi. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distributi