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| 003 | MX-MdCICY | ||
| 005 | 20250625160153.0 | ||
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| 090 | _aB-16538 | ||
| 245 | 1 | 0 | _aTranscriptome-wide target profiling of RNA cytosine methyltransferases using the mechanism-based enrichment procedure Aza-IP |
| 490 | 0 | _vNature Protocols, 9(2), p.337-361, 2014 | |
| 520 | 3 | _aCytosine methylation within RNA is common, but its full scope and functions are poorly understood, as the RNA targets of most mammalian cytosine RNA methyltransferases (m 5 C-RMTs)remain uncharacterized. To enable their characterization, we developed a mechanism-based method for transcriptome-wide m 5 C-RMT target profiling. All characterized mammalian m 5 C-RMTs form a reversible covalent intermediate with their cytosine substrate-a covalent linkage that is trapped when conducted on the cytosine analog 5-azacytidine (5-aza-C). We used this property to develop Aza-immunoprecipitation (Aza-IP), a methodology to form stable m 5 C-RMT-RNA linkages in cell culture, followed by IP and high-throughput sequencing, to identify direct RNA substrates of m 5 C-RMTs. Remarkably, a cytosine-to-guanine (C?G)transversion occurs specifically at target cytosines, allowing the simultaneous identification of the precise target cytosine within each RNA. Thus, Aza-IP reports only direct RNA substrates and the C?G transversion provides an important criterion for target cytosine identification, which is not available in alternative approaches. Here we present a step-by-step protocol for Aza-IP and downstream analysis, designed to reveal identification of substrate RNAs and precise cytosine targets of m 5 C-RMTs. The entire protocol takes 40-50 d to complete. © 2014 Nature America, Inc. All rights reserved. | |
| 650 | 1 | 4 | _aAZACITIDINE |
| 650 | 1 | 4 | _aCYTOSINE |
| 650 | 1 | 4 | _aGENE EXPRESSION PROFILING |
| 650 | 1 | 4 | _aHIGH-THROUGHPUT NUCLEOTIDE SEQUENCING |
| 650 | 1 | 4 | _aHUMANS |
| 650 | 1 | 4 | _aIMMUNOPRECIPITATION |
| 650 | 1 | 4 | _aTRNA METHYLTRANSFERASES |
| 700 | 1 | 2 | _aKhoddami, V. |
| 700 | 1 | 2 | _aCairns, B.R. |
| 856 | 4 | 0 |
_uhttps://drive.google.com/file/d/13F9rvYwG6vF8-8UZqI_Y0ma5isnDV4Ok/view?usp=drivesdk _zPara ver el documento ingresa a Google con tu cuenta: @cicy.edu.mx |
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