Nucleotide resolution profiling of m⁷G tRNA modification by TRAC-Seq.

ABSTRACT: Precise identification of sites of RNA modification is key to studying the functional role of such modifications in the regulation of gene expression and for elucidating relevance to diverse physiological processes. tRNA reduction and cleavage sequencing (TRAC-Seq) is a chemically based approach for the unbiased global mapping of 7-methylguansine (m⁷G) modification of tRNAs at single-nucleotide resolution throughout the tRNA transcriptome. m⁷G TRAC-Seq involves the treatment of size-selected (<200 nt) RNAs with the demethylase AlkB to remove major tRNA modifications, followed by sodium borohydride (NaBH₄) reduction of m⁷G sites and subsequent aniline-mediated cleavage of the RNA chain at the resulting abasic sites. The cleaved sites are subsequen

SUBMITTER: Lin S 

PROVIDER: S-EPMC8959837 | biostudies-literature | 2019 Nov

REPOSITORIES: biostudies-literature