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A programmable system to methylate and demethylate N6-methyladenosine (m6A) on specific RNA transcripts in mammalian cells
Chang, Chen1,2; Ma, Gang1,3; Cheung, Edwin2; Hutchins, Andrew P.1,3
2022-09-23
Source PublicationJOURNAL OF BIOLOGICAL CHEMISTRY
ISSN1083-351X
Volume298Issue:11Pages:102525
Abstract

RNA N-methyladenosine (mA) is the most abundant internal mRNA modification and forms part of an epitranscriptomic system that modulates RNA function. mA is reversibly catalyzed by specific enzymes, and those modifications can be recognized by RNA-binding proteins that in turn regulate biological processes. Although there are many reports demonstrating mA participation in critical biological functions, this exploration has mainly been conducted through the global KO or knockdown of the writers, erasers, or readers of mA. Consequently, there is a lack of information about the role of mA on single transcripts in biological processes, posing a challenge in understanding the biological functions of mA. Here, we demonstrate a CRISPR/dCas13a-based RNA mA editors, which can target RNAs using a single or multiple CRISPR RNA array to methylate or demethylate mA in human 293T cells and mouse embryonic stem cells. We systematically assay its capabilities to enable the targeted rewriting of mA dynamics, including modulation of circular RNA translation and transcript half-life. Finally, we use the system to specifically modulate mA levels on the noncoding XIST (X-inactive specific transcript) to modulate X chromosome silencing and activation. The editors described here can be used to explore the roles of mA in biological processes.

KeywordCrispr/cas13a Epitranscriptomics M6a X-chromosome Inactivation
DOI10.1016/j.jbc.2022.102525
URLView the original
Indexed BySCIE
Language英語English
WOS Research AreaBiochemistry & Molecular Biology
WOS SubjectBiochemistry & Molecular Biology
WOS IDWOS:000882627400009
PublisherELSEVIERRADARWEG 29, 1043 NX AMSTERDAM, NETHERLANDS
Scopus ID2-s2.0-85140208875
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Citation statistics
Document TypeJournal article
CollectionCancer Centre
Faculty of Health Sciences
Corresponding AuthorCheung, Edwin; Hutchins, Andrew P.
Affiliation1.Department of Biology, School of Life Sciences, Southern University of Science and Technology, Shenzhen, China
2.Cancer Centre, Faculty of Health Sciences, University of Macau, Taipa, China
3.Shenzhen Key Laboratory of Gene Regulation and Systems Biology, School of Life Sciences, Southern University of Science and Technology, Shenzhen, China
First Author AffilicationCancer Centre
Corresponding Author AffilicationCancer Centre
Recommended Citation
GB/T 7714
Chang, Chen,Ma, Gang,Cheung, Edwin,et al. A programmable system to methylate and demethylate N6-methyladenosine (m6A) on specific RNA transcripts in mammalian cells[J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2022, 298(11), 102525.
APA Chang, Chen., Ma, Gang., Cheung, Edwin., & Hutchins, Andrew P. (2022). A programmable system to methylate and demethylate N6-methyladenosine (m6A) on specific RNA transcripts in mammalian cells. JOURNAL OF BIOLOGICAL CHEMISTRY, 298(11), 102525.
MLA Chang, Chen,et al."A programmable system to methylate and demethylate N6-methyladenosine (m6A) on specific RNA transcripts in mammalian cells".JOURNAL OF BIOLOGICAL CHEMISTRY 298.11(2022):102525.
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