ONT-lrRNA-seq

In 1958, Francis Crick, the discoverer of the DNA double helix structure, put forward the “Central Dogma”, which elaborated on the information flow among DNA, RNA, and proteins. Gene transcription generates precursor RNA, which undergoes alternative splicing to form multiple transcripts, and these transcripts are then translated into different protein isoforms. Alternative splicing enables genes to produce multiple protein isoforms, which may have significant differences in structure and function and are specifically expressed in specific diseases, tissues, or developmental stages. Researchers have discussed the role of protein isoforms in drug discovery and how to utilize them to enhance drug specificity and therapeutic effects in the journal “Nature Reviews Drug Discovery” [1]. Therefore, studying the regulatory mechanisms of alternative splicing of precursor RNA and how these transcripts are translated into proteins and perform their functions is crucial for the development of treatments centered around protein isoforms. 

In the human genome, more than 90% of genes undergo alternative splicing to produce multiple transcripts. In short-read cDNA sequencing, the reads are often aligned to common exons, making it difficult to determine their origin. Although junction reads that span exons can improve the accuracy of analysis, their effectiveness is limited when transcriptional isoforms share splicing sites. ONT-IrRNA-seq (Oxford Nanopore Technologies long-read RNA sequencing) can directly detect full-length isoforms, capture unique transcripts, eliminate or reduce detection biases, and improve the accuracy of the analysis of differentially expressed transcripts.