Article ; Online: Global mapping of RNA G-quadruplexes (G4-RNAs) using G4RP-seq.
2022 Volume 17, Issue 3, Page(s) 870–889
Abstract: Guanine-rich RNAs can fold into four-stranded structures, termed G-quadruplexes (G4-RNAs), and participate in a wide range of biological processes. Here we describe in detail a G4-RNA-specific precipitation (G4RP) protocol, which enables the ... ...
Abstract | Guanine-rich RNAs can fold into four-stranded structures, termed G-quadruplexes (G4-RNAs), and participate in a wide range of biological processes. Here we describe in detail a G4-RNA-specific precipitation (G4RP) protocol, which enables the transcriptomic profiling of G4-RNAs. The G4RP protocol consists of a chemical cross-linking step, followed by affinity capture with a G4-specific probe, BioTASQ. G4RP can be coupled with sequencing to capture a comprehensive global snapshot of folded G4-RNAs. This method can also be used to profile induced changes (i.e., through G4 ligand treatments) within the G4-RNA transcriptome. The entire protocol can be completed in 1-2 weeks and can be scaled up or down depending on the specific experimental goals. In addition to the protocol details, we also provide here a guide for optimization in different laboratory setups. |
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MeSH term(s) | G-Quadruplexes ; Ligands ; RNA/chemistry ; RNA/genetics ; Transcriptome |
Chemical Substances | Ligands ; RNA (63231-63-0) |
Language | English |
Publishing date | 2022-02-09 |
Publishing country | England |
Document type | Journal Article ; Research Support, Non-U.S. Gov't ; Review |
ZDB-ID | 2244966-8 |
ISSN | 1750-2799 ; 1754-2189 |
ISSN (online) | 1750-2799 |
ISSN | 1754-2189 |
DOI | 10.1038/s41596-021-00671-6 |
Database | MEDical Literature Analysis and Retrieval System OnLINE |
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