Deep sequencing of yeast and mouse tRNAs and tRNA fragments using OTTR.

ABSTRACT: Among the major classes of RNAs in the cell, tRNAs remain the most difficult to characterize via deep sequencing approaches, as tRNA structure and nucleotide modifications can each interfere with cDNA synthesis by commonly used reverse transcriptases (RTs). Here, we benchmark a recently developed RNA cloning protocol, termed Ordered Two-Template Relay (OTTR), to characterize intact tRNAs and tRNA fragments in budding yeast and in mouse tissues. We show that OTTR successfully captures both full-length tRNAs and tRNA fragments in budding yeast and in mouse reproductive tissues without any prior enzymatic treatment, and that tRNA cloning efficiency can be further enhanced via AlkB-mediated demethylation of modified nucleotides. As with other recent tRNA cloning protocols, we find that a subse

SUBMITTER: Gustafsson HT 

PROVIDER: S-EPMC12140624 | biostudies-literature | 2025 Apr

REPOSITORIES: biostudies-literature