Fadia Ibrahim, PhD, & colleagues develop a native RNA sequencing technique (in Nucleic Acids Research)

To understand mRNA decay at single-molecule resolution, we developed True End-to-end RNA Sequencing (TERA-Seq) for Oxford Nanopore Technology platform. TERA-Seq describes both poly- and non-polyadenylated RNA molecules and accurately identifies their native 5′ and 3′ ends by ligation of unique adapters. TERA-Seq permits thorough transcriptome characterization and reveals novel insights into human mRNA decay. With TERA-Seq, we report RNA processing and decay at single-molecule level and find that mRNAs decay co-translationally, often from their 5′ ends. We reveal prevalent capping downstream of canonical transcriptional start sites in otherwise fully spliced and polyadenylated molecules. We also find that mRNA decay is not strictly dependent upon prior deadenylation challenging the prevailing view that eukaryotic mRNA decay initiates with poly(A) tail removal. TERA-Seq will be eminently suitable for applications where true end-to-end direct sequencing of single, native RNA molecules and their modifications is desired.