RNA结合蛋白与RNA相互作用鉴定技术
TechniquePrincipleAdvantageDisadvantageApplicationRIP⁃ChipImmunoprecipitationoraffinityprecipitationwithamicroarrayanal⁃ysisDisplayinginformationonRNA⁃proteinbind⁃inginnearnativecon⁃ditionNon⁃identificationofexactcrosslinkingsitesIsolationandidentificationofRBPcomplexesCLIP，HITS⁃CLIP254nmUVcrosslink⁃ing，combinedwithhigh⁃throughputsequencingMappingRNA⁃proteininteractionsonage⁃nomicscale，identifyingex⁃actcrosslinksitesandachievingindividualnucleotideresolutionLimitedbyUVpenetrability，prefer⁃ence，efficiencyofcrosslinkingMultiplebiologicalsamplesPAR⁃CLIP>320nmUVcrosslinking，incorpo⁃rationofphotoactivat⁃ablenucleotideMarkingbindingsitewithacharacteristicmutation(T⁃to⁃Cwhenusing4SU，G⁃to⁃Awhenusing6SG)Cytotoxicityofphotoactivatableri⁃bonucleosideCulturecellsiCLIPCrosslinksitescausingprematuretruncationofreversetranscriptaseEliminatingpreferenceofPCRamplificationandincompletedigestionofproteaseKMultipleexperimen⁃talprocedureHigher⁃orderribo⁃nucleoproteincom⁃plexesTRIBEExpressionofRBP⁃dADARfusionproteininvivoMarkingbindingsitewithnovelRNAeditingevents，requiringnobiochemistryRequiringanedit⁃ablesubstrateproximaltothebindingsiteanddouble⁃strandedRNAMultipleRBPsandcell⁃specificRBPtargetsRNAtaggingExpressionofRBP⁃po⁃ly(U)polymerasefu⁃sionproteininvivoMarkingtargetsbyaddinguridines，re⁃quiringnobiochemis⁃tryandminimalstart⁃ingmaterialNon⁃identificationofexactcrosslinkingsitesNon⁃canonicalRB⁃Psandcell⁃specificRBPtargetsIC，SerICHybridizingwithaffini⁃ty⁃taggedoligod(T)HighprecisionNon⁃identificationofexactcrosslinkingsitesPoly(A)RNAs⁃bindingproteome
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