Selective Enrichment of A-to-I Edited Transcripts from Cellular RNA Using Endonuclease V.

ABSTRACT: Creating accurate maps of A-to-I RNA editing activity is vital to improving our understanding of the biological role of this process and harnessing it as a signal for disease diagnosis. Current RNA sequencing techniques are susceptible to random sampling limitations due to the complexity of the transcriptome and require large amounts of RNA material, specialized instrumentation, and high read counts to accurately interrogate A-to-I editing sites. To address these challenges, we show that Escherichia coli Endonuclease V (eEndoV), an inosine-cleaving enzyme, can be repurposed to bind and isolate A-to-I edited transcripts from cellular RNA. While Mg²⁺ enables eEndoV to catalyze RNA cleavage, we show that similar levels of Ca²⁺ instead promote binding of inosine wi

SUBMITTER: Knutson SD 

PROVIDER: S-EPMC7286354 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature