Nanopore direct RNA sequencing detects differential expression between human cell populations

A crucial requirement of RNA sequencing is identifying differential expression to understand cell functions, differentiation and disease. The development of long-read direct RNA (dRNA) sequencing may overcome many limitations of short-read sequencing. Our study investigates the ability of dRNA sequencing to identify differential gene and isoform expression in both synthetic controls and human cells. We show that dRNA sequencing accurately quantifies RNA and identifies differential expression between conditions. We further identified thousands of novel transcripts and splice isoforms. Our results establish the ability of dRNA sequencing to identify biologically relevant differences in gene and isoform expression and perform the key capabilities of expression profiling methodologies.