Format

Send to

Choose Destination
Nat Methods. 2019 Sep 23. doi: 10.1038/s41592-019-0570-0. [Epub ahead of print]

DART-seq: an antibody-free method for global m6A detection.

Author information

1
Department of Biochemistry, Duke University School of Medicine, Durham, NC, USA. kate.meyer@duke.edu.
2
Department of Neurobiology, Duke University School of Medicine, Durham, NC, USA. kate.meyer@duke.edu.

Abstract

N6-methyladenosine (m6A) is a widespread RNA modification that influences nearly every aspect of the messenger RNA lifecycle. Our understanding of m6A has been facilitated by the development of global m6A mapping methods, which use antibodies to immunoprecipitate methylated RNA. However, these methods have several limitations, including high input RNA requirements and cross-reactivity to other RNA modifications. Here, we present DART-seq (deamination adjacent to RNA modification targets), an antibody-free method for detecting m6A sites. In DART-seq, the cytidine deaminase APOBEC1 is fused to the m6A-binding YTH domain. APOBEC1-YTH expression in cells induces C-to-U deamination at sites adjacent to m6A residues, which are detected using standard RNA-seq. DART-seq identifies thousands of m6A sites in cells from as little as 10‚ÄČng of total RNA and can detect m6A accumulation in cells over time. Additionally, we use long-read DART-seq to gain insights into m6A distribution along the length of individual transcripts.

PMID:
31548708
DOI:
10.1038/s41592-019-0570-0

Supplemental Content

Full text links

Icon for Nature Publishing Group
Loading ...
Support Center