Abstract
While splicing changes caused by somatic mutations in SF3B1 are known, identifying fulllength isoform changes may better elucidate the functional consequences of these mutations.
We report nanopore sequencing of full-length cDNA from CLL samples with and withoutSF3B1 mutation, as well as normal B cell samples, giving a total of 149 million pass reads. We present FLAIR (Full-Length Alternative Isoform analysis of RNA), a computational workflow to identify high-confidence transcripts, perform differential splicing event analysis, and differential isoform analysis. Using nanopore reads, we demonstrate differential 3’ splice site changes associated with SF3B1 mutation, agreeing with previous studies. We also observe a strong downregulation of intron retention events associated with SF3B1 mutation. Full-length transcript analysis links multiple alternative splicing events together and allows for better estimates of the abundance of productive versus unproductive isoforms. Our work demonstrates the potential utility of nanopore sequencing for cancer and splicing research. Fig 1 Intron retention events are more strongly downregulated in CLL SF3B1K700E
 
References
Tang AD. et al. Full-length transcript characterization of SF3B1 mutation in chronic lymphocytic leukemia reveals downregulation of retained introns. Nature Communications,2020