Genome-wide detection of high abundance N6-methyladenosine sites by microarray |
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| Authors: | Yue Li Yang Wang Zhaolei Zhang Alicia Viridiana Zamudio Jing Crystal Zhao |
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| Affiliation: | 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 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 |
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| Abstract: | N6-methyladenosine (m6A), 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 m6A in the transcriptome rely on the availability and quality of an m6A antibody and are often associated with a high rate of false positives. Here, based on our observation that m6A interferes with A–T/U pairing, we report a microarray-based technology to map m6A sites in mouse embryonic stem cells. We identified 72 unbiased sites exhibiting high m6A 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 m6A sites in the mammalian transcriptome. This method provides an alternative means to identify m6A sites for certain applications. |
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| Keywords: | N6-methyladenosine two-color microarray RNA methylation METTL3 METTL14 mouse embryonic stem cells |
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