Analysis of RNA conformation in endogenously assembled RNPs by icSHAPE

Analysis of RNA conformation in endogenously assembled RNPs by icSHAPE

Summary: The majority of the mammalian genome is transcribed into non-coding RNAs, many of which co-evolve with RNA-binding proteins (RBPs) to function as biochemically defined and tractable ribonucleoproteins (RNPs). Here, we applied icSHAPE- a robust and versatile RNA structural probing pipeline-...

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Bibliographic Details
Main Authors: Lu Chen, Howard Y. Chang, Steven E. Artandi
Format: Article
Language:English
Published: Elsevier 2021-06-01
Series:STAR Protocols
Subjects:
Cell separation/fractionation
Sequencing
RNA-seq
Molecular Biology
Gene Expression
CRISPR
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166721001842
Description
Summary:Summary: The majority of the mammalian genome is transcribed into non-coding RNAs, many of which co-evolve with RNA-binding proteins (RBPs) to function as biochemically defined and tractable ribonucleoproteins (RNPs). Here, we applied icSHAPE- a robust and versatile RNA structural probing pipeline- to endogenous RNPs purified from nuclei, providing base-resolution structural rationale for RNP activity and subcellular localization. Combining with genetic and biochemical reconstitutions, structural and functional alternations can be directly attributed to a given RBP without ambiguity.For complete details on the use and execution of this protocol, please refer to Chen et al. (2018).
ISSN:2666-1667

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