Genome-Wide Interrogation of Mammalian Stem Cell Fate Determinants by Nested Chromosome Deletions 英文参考文献.docVIP

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Genome-Wide Interrogation of Mammalian Stem Cell Fate Determinants by Nested Chromosome Deletions 英文参考文献.doc

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Genome-Wide Interrogation of Mammalian Stem Cell Fate Determinants by Nested Chromosome Deletions 英文参考文献

Genome-WideInterrogationofMammalianStemCell FateDeterminantsbyNestedChromosomeDeletions SimonFortier1.,Me′lanieBilodeau1.,TaraMacRae1,Jean-PhilippeLaverdure1,ValeriaAzcoitia1, SimonGirard1,JalilaChagraoui1,NancyRinguette1,Jose′eHe′bert1,2,JanaKrosl1,NadineMayotte1, GuySauvageau1,2 * 1MolecularGeneticsofStemCellsLaboratory,InstituteofResearchinImmunologyandCancer(IRIC),UniversityofMontreal,Montreal,Quebec,Canada,2Divisionof HematologyandLeukemiaCellBankofQuebec(BCLQ),Maisonneuve-RosemontHospital,Montreal,Quebec,Canada Abstract Understanding the function of important DNA elements in mammalian stem cell genomes would be enhanced by the availability of deletion collections in which segmental haploidies are precisely characterized. Using a modified Cre-loxP– basedsystem,wenowreportthecreationandcharacterizationofacollectionof,1,300independentembryonicstemcell (ESC)clonesenrichedfornestedchromosomaldeletions.Mappingexperimentsindicatethatthiscollectionspansover25% ofthemousegenomewithgoodrepresentativecoverageofprotein-codinggenes,regulatoryRNAs,andothernon-coding sequences.ThiscollectionofcloneswasscreenedforinvitrodefectsindifferentiationofESCintoembryoidbodies(EB). Severalputativenovelhaploinsufficientregions,criticalforEBdevelopment,wereidentified.Functionalcharacterizationof one of these regions, through BAC complementation, identified the ribosomal gene Rps14 as a novel haploinsufficient determinantofembryoidbodyformation.ThisnewlibraryofchromosomaldeletionsinESC(DelES:http://bioinfo.iric.ca/ deles)willserveasauniqueresourceforelucidationofnovelprotein-codingandnon-codingregulatorsofESCactivity. Citation: Fortier S,Bilodeau M, MacRaeT,LaverdureJ-P,AzcoitiaV,et al.(2010) Genome-WideInterrogation ofMammalianStem CellFateDeterminants by NestedChromosomeDeletions.PLoSGenet6(12):e1001241.doi:10.1371/journal.pgen.1001241 Editor:WayneN.Frankel,TheJacksonLaboratory,UnitedStatesofAmerica ReceivedDecember17,2009;AcceptedNovember5,2010;PublishedDecember9,2010 Copyright: