Long-Read Sequencing Reveals RNA Splicing Complexity in Human Diseases

Area of Science:

• Genomics
• Molecular Biology
• Bioinformatics

Background:

• Short-read RNA sequencing faces challenges in analyzing full-length RNA due to fragmented reads.
• Accurate characterization of alternative splicing, exon structures, and transcription start/termination sites is difficult with short reads.

Purpose of the Study:

• To provide an overview of long-read RNA sequencing (lrRNA-seq) principles, innovations, and advantages.
• To assess methodological aspects and applications of lrRNA-seq in transcriptomic research and disease studies.

Main Methods:

• Review of long-read RNA sequencing (lrRNA-seq) technology and its working principles.
• Systematic assessment of methods for isoform analysis, quantification, error correction, and algorithm development for lrRNA-seq data.

Main Results:

• lrRNA-seq enables end-to-end transcript sequencing, offering comprehensive insights into transcriptomic complexity.
• It precisely determines exon-intron structures, alternative splicing, transcription initiation/termination sites, and noncanonical RNA processing events.
• Applications in disease research reveal splicing dysregulation, pathogenic isoforms, and RNA-mediated mechanisms.

Conclusions:

• lrRNA-seq is a transformative technology for detailed transcriptome analysis, surpassing short-read limitations.
• Integration with single-cell and spatial transcriptomics allows characterization of splicing complexity across cellular and tissue microenvironments.
• lrRNA-seq holds significant potential for advancing disease diagnostics and precision medicine.