Transcriptome-Wide Detection of Internal N7-Methylguanosine

Li-Sheng Zhang; Chang Liu; Chuan He

doi:10.1007/978-1-0716-1374-0_6

Transcriptome-Wide Detection of Internal N⁷-Methylguanosine

Methods Mol Biol. 2021:2298:97-104. doi: 10.1007/978-1-0716-1374-0_6.

Authors

Li-Sheng Zhang^{1

2}, Chang Liu^{1

2}, Chuan He^{3

4

5}

Affiliations

¹ Department of Chemistry, The University of Chicago, Chicago, IL, USA.
² Howard Hughes Medical Institute, The University of Chicago, Chicago, IL, USA.
³ Department of Chemistry, The University of Chicago, Chicago, IL, USA. chuanhe@uchicago.edu.
⁴ Howard Hughes Medical Institute, The University of Chicago, Chicago, IL, USA. chuanhe@uchicago.edu.
⁵ Department of Biochemistry and Molecular Biology, Institute for Biophysical Dynamics, The University of Chicago, Chicago, IL, USA. chuanhe@uchicago.edu.

PMID: 34085240
DOI: 10.1007/978-1-0716-1374-0_6

Abstract

m⁷G-seq detects internal 7-methylguanosine (m⁷G) sites within mRNAs and noncoding RNAs by misincorporation signatures. A chemical-assisted sequencing approach selectively converts internal m⁷G sites into abasic sites, triggering misincorporation at these sites in the presence of a specific reverse transcriptase. The further enrichment of m⁷G-induced abasic sites by biotin pull-down reveals hundreds of internal m⁷G sites in human mRNA. The misincorporation ratio before pull-down enrichment can be used for estimating the methylation fraction of some highly methylated m⁷G sites.

Keywords: 7-Methylguanosine; Misincorporation; RNA epitranscriptomics; m7G-seq; mRNA methylation.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Guanosine / analogs & derivatives*
Guanosine / genetics
Humans
Methylation
RNA Processing, Post-Transcriptional / genetics
RNA, Messenger / genetics
Transcriptome / genetics*

Substances

RNA, Messenger
Guanosine
7-methylguanosine

Grants and funding

HHMI/Howard Hughes Medical Institute/United States