CIGAR-seq, a CRISPR/Cas-based method for unbiased screening of novel mRNA modification regulators.

ABSTRACT: Cellular RNA is decorated with over 170 types of chemical modifications. Many modifications in mRNA, including m⁶ A and m⁵ C, have been associated with critical cellular functions under physiological and/or pathological conditions. To understand the biological functions of these modifications, it is vital to identify the regulators that modulate the modification rate. However, a high-throughput method for unbiased screening of these regulators is so far lacking. Here, we report such a method combining pooled CRISPR screen and reporters with RNA modification readout, termed CRISPR integrated gRNA and reporter sequencing (CIGAR-seq). Using CIGAR-seq, we discovered NSUN6 as a novel mRNA m⁵ C methyltransferase. Subsequent mRNA bisulfite sequencing in HAP1 cells

SUBMITTER: Fang L 

PROVIDER: S-EPMC7701898 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature