Transcriptome-Wide Mapping of N6-Methyladenosine by m6A-Seq

Dan Dominissini; Sharon Moshitch-Moshkovitz; Ninette Amariglio; Gideon Rechavi

doi:10.1016/bs.mie.2015.03.001

Transcriptome-Wide Mapping of N⁶-Methyladenosine by m⁶A-Seq

Dan Dominissini
, Sharon Moshitch-Moshkovitz
, Ninette Amariglio
, Gideon Rechavi

The University of Chicago
Sheba Medical Center at Tel Hashomer
Tel Aviv University

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

29 Scopus citations

Abstract

A detailed protocol for isolation and sequencing of an enriched population of m⁶A-methylated RNA fragments to create m⁶A methylome maps is outlined. Our approach was developed to fill a void that existed because of a lack of methods for the detection of m⁶A in RNA in an unbiased, high-throughput, and high-resolution manner. This method integrates immunoprecipitation of methylated, randomly fragmented RNA using a highly specific anti-m⁶A antibody to obtain an enriched population of modified fragments and massively parallel sequencing, resulting in mapping of this modification throughout the transcriptome.

Original language	English
Title of host publication	Methods in Enzymology
Publisher	Academic Press Inc.
Pages	131-147
Number of pages	17
DOIs	https://doi.org/10.1016/bs.mie.2015.03.001
State	Published - 11 Aug 2015
Externally published	Yes

Publication series

Name	Methods in Enzymology
Volume	560
ISSN (Print)	0076-6879
ISSN (Electronic)	1557-7988

Bibliographical note

Publisher Copyright:
© 2015 Elsevier Inc. All rights reserved.

Funding

D.D. is supported by Human Frontier Science Program Long-Term Fellowship. G.R. is supported by grants from the Flight Attendant Medical Research Institute (FAMRI), Israel Science Foundation (Grant no. 1667/12), by the I-CORE Program and The Israel Science Foundation (Grant Nos. 41/11 and 1796/12), by Teva National Network of Excellence in Neuroscience (NNE), and by the Ernest and Bonnie Beutler Research Program. G.R. is a member of the Sagol Neuroscience Network and holds the Djerassi Chair for Oncology (Sackler Faculty of Medicine, Tel Aviv University, Israel).

Funders	Funder number
NNE
Teva National Network of Excellence in Neuroscience
Human Frontier Science Program
Flight Attendant Medical Research Institute
Israel Science Foundation	1667/12
Israeli Centers for Research Excellence	41/11, 1796/12

Keywords

Immunocapturing
N-Methyladenosine
Next-generation sequencing
RNA immunoprecipitation
RNA methylation

Access to Document

10.1016/bs.mie.2015.03.001

Cite this

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