Detection of internal N7-methylguanosine (m7G) RNA modifications by mutational profiling sequencing.

ABSTRACT: Methylation of guanosine on position N7 (m7G) on internal RNA positions has been found in all domains of life and have been implicated in human disease. Here, we present m7G Mutational Profiling sequencing (m7G-MaP-seq), which allows high throughput detection of m7G modifications at nucleotide resolution. In our method, m7G modified positions are converted to abasic sites by reduction with sodium borohydride, directly recorded as cDNA mutations through reverse transcription and sequenced. We detect positions with increased mutation rates in the reduced and control samples taking the possibility of sequencing/alignment error into account and use replicates to calculate statistical significance based on log likelihood ratio tests. We show that m7G-MaP-seq efficiently detects known m7G modifi

SUBMITTER: Enroth C 

PROVIDER: S-EPMC6847341 | biostudies-literature | 2019 Nov

REPOSITORIES: biostudies-literature