Capture NanoporeSeq: A novel technique for targeted full-length transcript sequencing and gene expression analysis (360G-Wellcome-201879_Z_16_Z)

The accurate characterization and quantification of expressed gene isoforms is essential to understanding how genetic variation affects gene expression and ultimately disease risk. However, the expressed isoforms identified by standard RNA sequencing methods are often incorrect due to the near ubiquitous presence of alternative gene isoforms in the human transcriptome. This project aims to address this problem by developing a novel technique (Capture NanoporeSeq) to perform full-length cDNA sequencing of target human genes and so unambiguously identify which isoforms are expressed. Capture NanoporeSeq will combine Oxford Nanopore long-read sequencing, which can sequence full-length cDNAs, with targeted RNA sequencing (CaptureSeq), which will provide high sensitivity for targeted gene isoforms. Once developed, Capture NanoporeSeq will be validated in post-mortem human brain tissue to identify and quantitate the expression of gene isoforms from genomic regions linked to neuropsychiatric disorders. Capture NanoporeSeq will significantly improve our ability to accurately characterize and quantify gene expression. This will be of immediate use in neuropsychiatric disease genetics where many risk genes have lowly expressed disease-linked isoforms and/or complex splicing patterns, helping to illuminate the impact that disease-associated genetic variation has on gene expression.