Direct long-read RNA sequencing identifies a subset of questionable exitrons likely arising from reverse transcription artifacts

Autor: Laura Schulz, Manuel Torres-Diz, Mariela Cortés-López, Katharina E. Hayer, Mukta Asnani, Sarah K. Tasian, Yoseph Barash, Elena Sotillo, Kathi Zarnack, Julian König, Andrei Thomas-Tikhonenko
Jazyk: angličtina
Rok vydání: 2021
Předmět:
Zdroj: Genome Biology, Vol 22, Iss 1, Pp 1-12 (2021)
Druh dokumentu: article
ISSN: 1474-760X
DOI: 10.1186/s13059-021-02411-1
Popis: Abstract Resistance to CD19-directed immunotherapies in lymphoblastic leukemia has been attributed, among other factors, to several aberrant CD19 pre-mRNA splicing events, including recently reported excision of a cryptic intron embedded within CD19 exon 2. While “exitrons” are known to exist in hundreds of human transcripts, we discovered, using reporter assays and direct long-read RNA sequencing (dRNA-seq), that the CD19 exitron is an artifact of reverse transcription. Extending our analysis to publicly available datasets, we identified dozens of questionable exitrons, dubbed “falsitrons,” that appear only in cDNA-seq, but never in dRNA-seq. Our results highlight the importance of dRNA-seq for transcript isoform validation.
Databáze: Directory of Open Access Journals