Evolution of a reverse transcriptase to map N¹-methyladenosine in human messenger RNA.

ABSTRACT: Chemical modifications to messenger RNA are increasingly recognized as a critical regulatory layer in the flow of genetic information, but quantitative tools to monitor RNA modifications in a whole-transcriptome and site-specific manner are lacking. Here we describe a versatile platform for directed evolution that rapidly selects for reverse transcriptases that install mutations at sites of a given type of RNA modification during reverse transcription, allowing for site-specific identification of the modification. To develop and validate the platform, we evolved the HIV-1 reverse transcriptase against N¹-methyladenosine (m¹A). Iterative rounds of selection yielded reverse transcriptases with both robust read-through and high mutation rates at m¹A sites. The

SUBMITTER: Zhou H 

PROVIDER: S-EPMC6884687 | biostudies-literature | 2019 Dec

REPOSITORIES: biostudies-literature