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

Abstract

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

Citations

David J Wright, Nicola Hall, Naomi Irish, Angela L Man, Will Glynn, Arne Mould, Alejandro De Los Angeles, Emily Angiolini, David Swarbreck, Karim Gharbi, Elizabeth M Tunbridge, Wilfried Haerty. Long read sequencing reveals novel isoforms and insights into splicing regulation during cell state changes. bioRxiv

Page last reviewed: 12 June, 2025

Metadata

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Collection: 123456789/464

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Date issued: 2021-04

ID: 846