Long read sequencing reveals novel isoforms and insights into splicing regulation during cell state changes

Abstract

'Will Glynn' 'Arne Mould' 'Alejandro De Los Angeles' 'Emily Angiolini' 'David Swarbreck' 'Karim Gharbi' 'Elizabeth M. Tunbridge' 'Wilfried Haerty'] Background Alternative splicing 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 alternative splicing 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 SH-SY5Y, and to characterise isoform expression and usage across differentiation. Results 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 identifying candidates for future research into state change regulation. Conclusions Our work highlights the potential of long read sequencing to uncover previously unknown transcript diversity and mechanisms influencing alternative splicing. Supplementary Information The online version contains supplementary material available at 10.1186/s12864-021-08261-2.