ABSTRACT
The steady state expression of each gene is the result of a dynamic transcription and degradation of that gene. While regular RNA-seq methods only measure steady state expression levels, RNA-seq of metabolically labeled RNA identifies transcripts that were transcribed during the window of metabolic labeling. Whereas short-read RNA sequencing can identify metabolically labeled RNA at the gene level, long-read sequencing provides much better resolution of isoform-level transcription. Here we combine thiouridine-to-cytosine conversion (TUC) with PacBio long-read sequencing to study the dynamics of mRNA transcription in the GM12878 cell line. We show that using long-TUC-seq, we can detect metabolically labeled mRNA of distinct isoforms more reliably than using short reads. Long-TUC-seq holds the promise of capturing isoform dynamics robustly and without the need for enrichment.
Competing Interest Statement
The authors have declared no competing interest.
Footnotes
Sorena Rahmanian: sorenar{at}uci.edu
Gabriela Balderrama-Gutierrez: gbalderr{at}uci.edu
Dana Wyman: dwyman{at}uci.edu
Cassandra Joan McGill: mcgillc{at}uci.edu
Kim Nguyen: knguyen4047{at}gmail.com
Robert Spitale: rspitale{at}uci.edu
Ali Mortazavi: ali.mortazavi{at}uci.edu