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Long-read RNA-seq demarcates cis- and trans-directed alternative RNA splicing

Giovanni Quinones-Valdez, Kofi Amoah, Xinshu Xiao

2025Nature Communications6 citationsDOIOpen Access PDF

Abstract

Genetic regulation of alternative splicing constitutes an important link between genetic variation and disease. Nonetheless, RNA splicing is regulated by both cis-acting elements and trans-acting splicing factors. Determining splicing events that are directed primarily by the cis- or trans-acting mechanisms will greatly inform our understanding of the genetic basis of disease. Here, we show that long-read RNA-seq, combined with our new method isoLASER, enables a clear segregation of cis- and trans-directed splicing events for individual samples. The genetic linkage of splicing is largely individual-specific, in stark contrast to the tissue-specific pattern of splicing profiles. Analysis of long-read RNA-seq data from human and mouse revealed thousands of cis-directed splicing events susceptible to genetic regulation. We highlight such events in the HLA genes whose analysis was challenging with short-read data. We also highlight novel cis-directed splicing events in Alzheimer’s disease-relevant genes such as MAPT and BIN1. Together, the clear demarcation of cis- and trans-directed splicing paves ways for future studies of the genetic basis of disease. Genetic variants can influence how RNA is spliced, shaping disease risk. Here, the authors present isoLASER, a stand-alone method using long-read RNA sequencing to distinguish cis- and trans-directed splicing, revealing new insights into genetic regulation of splicing.

Topics & Concepts

RNA splicingAlternative splicingBiologyGeneticsComputational biologyExonic splicing enhancerGeneSplicing factorMinigeneRNARNA-binding proteinGenomeIntronRibonucleoproteinHuman genomeHuman geneticsExonSpliceosomeRNA Research and SplicingRNA regulation and diseaseSingle-cell and spatial transcriptomics
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