Identification of high-confidence human poly(A) RNA isoform scaffolds using nanopore sequencing.

ABSTRACT: Nanopore sequencing devices read individual RNA strands directly. This facilitates identification of exon linkages and nucleotide modifications; however, using conventional direct RNA nanopore sequencing, the 5' and 3' ends of poly(A) RNA cannot be identified unambiguously. This is due in part to RNA degradation in vivo and in vitro that can obscure transcription start and end sites. In this study, we aimed to identify individual full-length human RNA isoforms among ∼4 million nanopore poly(A)-selected RNA reads. First, to identify RNA strands bearing 5' m⁷G caps, we exchanged the biological cap for a modified cap attached to a 45-nt oligomer. This oligomer adaptation method improved 5' end sequencing and ensured correct identification of the 5' m⁷G capped ends. Secon

SUBMITTER: Mulroney L 

PROVIDER: S-EPMC8906549 | biostudies-literature | 2022 Feb

REPOSITORIES: biostudies-literature