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1、中国科技论文在线http://www.paper.edu.cnConstructionofRecombinantLentivirusVectorTargetingBovineLXRαmRNAandItsSilencingEffectsonBovine#5MuscleSatelliteCells*LiuYongfeng,ZhaoLu,KuTing,YangXingbin(CollegeofFoodEngineeringandNutritionalScience,ShaanxiNormalUniversity,Xi'an710062)Abstract:T
2、heliverXreceptorα(LXRα)isamemberofthenuclearhormonereceptorsuperfamilywhichcouldregulatethetranscriptionofthegenesinvolvedincholesteroltransportation.Recent10studiesshowedthatLXRαisconsideredasacriticalregulatorincholesterolhomeostasisinmacrophages,whichcouldregulateseveralgene
3、sinvolvedincholesteroltransport,suchastheATP-bindingcassettetrans-porters(ABCs),ABCA1,ABCG1,apolipoproteinE(ApoE)andlipoproteinlipase(LPL).Inourstudy,fourpairsofinhibitionshRNAweredesignedtoexclusivelytargetbovineLXRαmRNA.LentiviralvectorcontainingLXRαshRNAswereconstructedthrou
4、ghBP15andLPrecombinationsystem,andusedtoobtainedthecorrespondinglentivirusesin293Tcells.ThetitersoftheLenti-14MR0054-01,-2,-3and-04viruseswere3×108TU/ml,2×108TU/ml,2.5×108TU/ml,3.5×108TU/ml,respectively.TheknockdownefficiencyofpLenti-03targetingLXRαreached88%inbovinemusclesatel
5、litecells.Furthermore,themRNAexpressionofRXRαwasupregulatedinbovinemusclesatellitecells,whereasthatofPPARα,PPARγ,ABCA1,LPL,ApoE20weredownregulatedat48hpost-pLenti-03virusesinfection.Therefore,theefficientlentivirusvectorinhibitingbovineLXRαexpressionwasobtainedinthepresentstudy
6、,providingbasisforloss-of-functionofLXRαinbovinemusclesatellitecellsKeywords:bovineLXRαgene;lentivirus;vectorconstruction;genesilencing250IntroductionObesity,hypertension,dysmetabolismarediseasescausedbyunhealthydietcustomwhichinfluencedhuman’slivingquality.Thus,alow-cholestero
7、llifestyleisnecessary.Currently,peoplehavepaidmuchattentiontobalancingtheirdietbylow-intakeofcholesterol.Producinggoodqualitymeatwithlow-fat(beef,mutton,pork,etc.)isavitalwaytoprovidepeoplewithhealthymealscondition.30TheliverXreceptorα(LXRα,NR1H3)andLXRβ(NR1H2)aremembersofthenu
8、clearhormonereceptorsuperfamily.LXRαisabundantlyexpres
