PT  - JOURNAL ARTICLE
AU  - David J Wright
AU  - Nicola Hall
AU  - Naomi Irish
AU  - Angela L Man
AU  - Will Glynn
AU  - Arne Mould
AU  - Alejandro De Los Angeles
AU  - Emily Angiolini
AU  - David Swarbreck
AU  - Karim Gharbi
AU  - Elizabeth M Tunbridge
AU  - Wilfried Haerty
TI  - Long read sequencing reveals novel isoforms and insights into splicing regulation during cell state changes
AID  - 10.1101/2021.04.27.441628
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.04.27.441628
4099  - http://biorxiv.org/content/early/2021/04/28/2021.04.27.441628.short
4100  - http://biorxiv.org/content/early/2021/04/28/2021.04.27.441628.full
AB  - Alternative splicing (AS) is a key mechanism underlying cellular differentiation and a driver of complexity in mammalian neuronal tissues. However, understanding of which isoforms are differentially used or expressed and how this affects cellular differentiation remains unclear. Long read sequencing allows full-length transcript recovery and quantification, enabling transcript-level analysis of AS processes and how these change with cell state. Here, we utilise Oxford Nanopore Technologies sequencing to produce a custom annotation of a well-studied human neuroblastoma cell line and to characterise isoform expression and usage across differentiation. We identify many previously unannotated features, including a novel transcript of the voltage-gated calcium channel subunit gene, CACNA2D2. We show differential expression and usage of transcripts during differentiation, and identify a putative molecular regulator underlying this state change. Our work highlights the potential of long read sequencing to uncover previously unknown transcript diversity and mechanisms influencing alternative splicing.Competing Interest StatementThe authors have declared no competing interest.