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

2022-11-01

10.1016/j.jbc.2022.102525

JOURNAL OF BIOLOGICAL CHEMISTRY   影响因子和分区

0021-9258

1083-351X

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.

CRISPR/Cas13a epitranscriptomics m6A X-chromosome inactivation

SCI

英语

NI论文

第一 ; 通讯

National Natural Science Foundation of China[31970589];National Natural Science Foundation of China[32150710521];Universidade de Macau[MYRG2020-00100-FHS];

Biochemistry & Molecular Biology

Biochemistry & Molecular Biology

WOS:000882627400009

ELSEVIER

BIOLOGY & BIOCHEMISTRY

2-s2.0-85140208875

Scopus

1.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

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).

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).

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).