Programmable m6A modification of cellular RNAs with a Cas13-directed methyltransferase.

Abstract:

:N6-Methyladenosine (m6A) is the most widespread internal messenger RNA modification in humans. Despite recent progress in understanding the biological roles of m6A, the inability to install m6A site specifically in individual transcripts has hampered efforts to elucidate causal relationships between the presence of a specific m6A and phenotypic outcomes. In the present study, we demonstrate that nucleus-localized dCas13 fusions with a truncated METTL3 methyltransferase domain and cytoplasm-localized fusions with a modified METTL3:METTL14 methyltransferase complex can direct site-specific m6A incorporation in distinct cellular compartments, with the former fusion protein having particularly low off-target activity. Independent cellular assays across multiple sites confirm that this targeted RNA methylation (TRM) system mediates efficient m6A installation in endogenous RNA transcripts with high specificity. Finally, we show that TRM can induce m6A-mediated changes to transcript abundance and alternative splicing. These findings establish TRM as a tool for targeted epitranscriptome engineering that can reveal the effect of individual m6A modifications and dissect their functional roles.

journal_name

Nat Biotechnol

journal_title

Nature biotechnology

authors

Wilson C,Chen PJ,Miao Z,Liu DR

doi

10.1038/s41587-020-0572-6

subject

Has Abstract

pub_date

2020-12-01 00:00:00

pages

1431-1440

issue

12

eissn

1087-0156

issn

1546-1696

pii

10.1038/s41587-020-0572-6

journal_volume

38

pub_type

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