2. Academic Validation
  3. Genome-wide detection of high abundance N6-methyladenosine sites by microarray

Genome-wide detection of high abundance N6-methyladenosine sites by microarray

  • RNA. 2015 Aug;21(8):1511-8. doi: 10.1261/rna.051474.115.
Yue Li 1,  Yang Wang 2,  Zhaolei Zhang 3,  Alicia Viridiana Zamudio 4,  Jing Crystal Zhao 2
Affiliations

Affiliations

  • 1 Department of Computer Science, University of Toronto, Toronto M5S 3G4, Canada.
  • 2 Tumor Initiation and Maintenance Program, Sanford Burnham Medical Research Institute, San Diego, California 92037, USA.
  • 3 Department of Computer Science, University of Toronto, Toronto M5S 3G4, Canada Department of Molecular Genetics, Donnelly Centre for Cellular and Biomolecular Research, Banting and Best Department of Medical Research, University of Toronto, Toronto M5S 3E1, Canada.
  • 4 Department of Biology, San Diego State University, San Diego, California, 92115, USA.
PMID: 26092943 DOI: 10.1261/rna.051474.115
Abstract

N(6)-methyladenosine (m(6)A), the most abundant internal RNA modification, functions in diverse biological processes, including regulation of embryonic stem cell self-renewal and differentiation. As yet, methods to detect m(6)A in the transcriptome rely on the availability and quality of an m(6)A antibody and are often associated with a high rate of false positives. Here, based on our observation that m(6)A interferes with A-T/U pairing, we report a microarray-based technology to map m(6)A sites in mouse embryonic stem cells. We identified 72 unbiased sites exhibiting high m(6)A levels from 66 PolyA RNAs. Bioinformatics analyses suggest identified sites are enriched on developmental regulators and may in some contexts modulate MicroRNA/mRNA interactions. Overall, we have developed microarray-based technology to capture highly enriched m(6)A sites in the mammalian transcriptome. This method provides an alternative means to identify m(6)A sites for certain applications.

Keywords

METTL14; METTL3; N6-methyladenosine; RNA methylation; mouse embryonic stem cells; two-color microarray.

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