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1、Articlestranscriptomeinvivoanalysis(tiVA)ofspatiallydefinedsinglecellsinlivetissueDitteLovatt1,6,BrittaniKRuble2,6,JaeheeLee1,HannahDueck3,TaeKyungKim1,StephenFisher3,ChantalFrancis3,JenniferMSpaethling1,JohnAWolf4,MSeanGrady4,AlexandraVUlyanova4,SeanBYeldell2,JulianneCGrie
2、penburg2,PeterTBuckley1,JunhyongKim3,5,Jai-YoonSul1,IvanJDmochowski2,7&JamesEberwine1,5,7transcriptomeprofilingofsinglecellsresidentintheirnaturalrelyonsortingcellsinsuspensionfromacutelydissociatedmicroenvironmentdependsuponrnAcapturemethodsthattissues,inwhichinformationab
3、outcellmorphologyandthearebothnoninvasiveandspatiallyprecise.Weengineeredmicroenvironmentislost,andwherecellvariabilityismaskedatranscriptomeinvivoanalysis(tiVA)tag,whichuponbytheaveragingeffect11.Othermethods,suchaslasercapturephotoactivationenablesmrnAcapturefromsinglecel
4、lsinlivemicrodissectionandpatch-pipetteaspiration12,13,canbeusedtotissue.usingthetiVAtagincombinationwithrnAsequencingisolatesinglecellsintissue,butbothapproacheshavelimitations,(rnA-seq),weanalyzedtranscriptomevarianceamongsingleincludingpotentialRNAcontaminationfromotherc
5、ellsininci-neuronsincultureandinmouseandhumantissueinvivo.dentalcontactwiththepatchpipette.Furthermore,theformerisourdatashowedthatthetissuemicroenvironmentshapesperformedonfixedtissue,andthelatterpromptsconcernaboutthetranscriptomiclandscapeofindividualcells.thetiVAtranscr
6、iptionalchangesassociatedwithmechanicalinjuryduringmethodologyis,toourknowledge,thefirstnoninvasivetheisolationofRNA14.Hence,anmRNAcapturemethodologyapproachforcapturingmrnAfromlivesinglecellsintheirwithhighspatialresolutionandthatiscompatiblewithlive,intactnaturalmicroenvi
7、ronment.tissuewouldbeausefultooltoexplorethetranscriptomesofsinglecellsintheirnaturalmicroenvironment.MulticellularorganismsarecomposedofamyriadofcellsthatHerewedescribeamethodforisolatingmRNAfromasingleNatureAmerica,Inc.Allrightsreserved.arecategorizedintodifferenttypesbas
8、edonphenotypictraits.cellincomplextissuesusingaphotoactivatablemRNAcapture4However
