胸腺上皮细胞中雌激素诱导lncRNA的鉴定分析及表达载体构建.docx

胸腺上皮细胞中雌激素诱导lncRNA的鉴定分析及表达载体构建
Title:IdentificationandAnalysisofEstrogen-InducedlncRNAsinThymicEpithelialCellsandConstructionofExpressionVectors
Abstract:
Thymicepithelialcells(TECs)playavitalroleinTcelldevelopmentandimmunetolerance.PreviousstudieshaveshowntheregulatoryroleofestrogeninTECs,buttheunderlyingmolecularmechanismsremainlargelyunknown.Longnon-codingRNAs(lncRNAs)haveemergedaskeyregulatorsofcellularprocesses,includingthoseinvolvedinimmuneresponses.Thisstudyaimstoidentifyandanalyzeestrogen-inducedlncRNAsinTECsandconstructsuitableexpressionvectorsfortheirfunctionalanalysis.
Introduction:
ThethymusisaprimarylymphoidorgancrucialforTcelldevelopmentandmaturation.Thymicepithelialcells,includingcorticalandmedullaryTECs,createamicroenvironmentnecessaryforproperTcelleducation.Estrogenhasbeenshowntoinfluencethymicdevelopmentandfunction,highlightingitsimportanceintheregulationofimmuneresponses.However,thespecificmechanismsbywhichestrogenaffectsTECfunctionremainpoorlyunderstood.RecentstudieshaveshownthatlncRNAsplaycriticalrolesinimmunecelldevelopment,activation,andtolerance.Therefore,investigatingestrogen-inducedlncRNAsinTECsmightprovidevaluableinsightsintotheregulatorymechanismsofestrogeninthymicfunction.
Methods:
1.TECIsolationandCulture:TECswillbeisolatedfrommousethymiusingenzymaticdigestionandimmunomagneticseparation.Thesecellswillthenbeculturedinvitrowithorwithoutestrogentreatmentforsubsequentanalysis.
2.RNASequencingandDataAnalysis:TotalRNAwillbeextractedfromestrogen-treatedanduntreatedTECs.RNAsequencingwillbeperformed,anddifferentialexpressionanalysiswillbeconductedtoidentifyestrogen-inducedlncRNAsinTECs.
3.ValidationoflncRNAExpression:Quantitativereal-timePCRwillbeusedtovalidatetheexpressionofcandidatelncRNAsidentifiedfromRNAsequencing.
4.FunctionalAnalysisoflncRNAs:Gain-of-functionandloss-of-functionexperimentswillbeconductedusingsuitablelncRNAexpressionvectorsandgeneknockouttechniquestoinvestigatethefunctionalrolesofestrogen-inducedlncRNAsinTECs.
5.Bioinformati