Analysis of Blood Stem Cell Activity and Cystatin Gene Expression in a Mouse Model Presenting a Chromosomal Deletion Encompassing Csta and Stfa2l1 英文参考文献.docVIP

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Analysis of Blood Stem Cell Activity and Cystatin Gene Expression in a Mouse Model Presenting a Chromosomal Deletion Encompassing Csta and Stfa2l1 英文参考文献.doc

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Analysis of Blood Stem Cell Activity and Cystatin Gene Expression in a Mouse Model Presenting a Chromosomal Deletion Encompassing Csta and Stfa2l1 英文参考文献

AnalysisofBloodStemCellActivityandCystatinGene ExpressioninaMouseModelPresentingaChromosomal DeletionEncompassingCstaandStfa2l1 Me′lanieBilodeau1,TaraMacRae1,LouisGaboury1,Jean-PhilippeLaverdure1,Marie-PierreHardy1, NadineMayotte1,Ve′roniqueParadis1,Se′bastienHarton1,ClaudePerreault1,2,3,GuySauvageau1,2,3 * 1Institute for Research in Immunology and Cancer, University of Montreal, Montreal, Quebec, Canada, 2Department of Medicine, University of Montreal, Montreal, Quebec,Canada,3LeukemiaCellBankofQuebecandDivisionofHematology,Maisonneuve-RosemontHospital,Montreal,Quebec,Canada Abstract Thecystatinproteinsuperfamilyis characterized bythepresenceofconserved sequencesthatdisplaycysteine protease inhibitoryactivity(e.g.,towardscathepsins).Type1and2cystatinsareencodedby25genesofwhich23aregroupedin2 clusterslocalizedonmousechromosomes16and2.Theexpressionandessentialrolesofmostofthesegenesinmouse developmentandhematopoiesisremainpoorlycharacterized.Inthisstudy,wedescribeasetofquantitativereal-timePCR assaysandaglobalexpressionprofileofcystatingenesinnormalmousetissues.BenefitingfromourcollectionofDelES embryonic stem cell clones harboring large chromosomal deletions (to be reported elsewhere), we selected a clone in whicha95-kbregionofchromosome16ismissing(Del16qB3D/+).Inthisparticularclone,2cystatingenes,namelyCstaand Stfa2l1 are absent along with 2 other genes (Fam162a, Ccdc58) and associated intergenic regions. From this line, we establishedanewhomozygousmutantmousemodel(Del16qB3D/16qB3D)toassesstheinvivobiologicalfunctionsofthe2 deletedcystatins.Stfa2l1geneexpressionishighinwild-typefetalliver,bonemarrow,andspleen,whileCstaisubiquitously expressed. Homozygous Del16qB3D/16qB3D animals are phenotypically normal, fertile, and not overtly susceptible to spontaneousorirradiation-inducedtumorformation.Thehematopoieticstemandprogenitorcellactivityinthesemutant mice are also normal. Interestingly, quantitative real-time PCR expression profiling reveals a marked increase