大多数临床诊断和基因组研究机构几乎只关注编码区和重要剪接位点，从而忽略了其他非编码变体。因此，可能促进包括癌症在内的一系列疾病的错误剪接事件的内含子变异尚未得到系统研究。此类研究需要基因组和转录组数据，但目前满足这些要求的数据集很少。我们通过开发一种单核全长 RNA 测序方法来解决这个问题，该方法可以检测潜在致病性内含子变异。我们通过应用胰腺癌肿瘤和肿瘤来源的标本并将内含子变异与剪接失调联系起来，证明了我们方法的效力。我们特别发现肿瘤共有显着的内含子保留和假外显子激活事件，并影响编码关键转录调节因子的基因。我们的工作为评估和利用内含子突变作为癌症中强大的预后标志物和潜在治疗靶点铺平了道路。© 作者 2024。由牛津大学出版社代表 NAR 基因组学和生物信息学出版。

Most clinical diagnostic and genomic research setups focus almost exclusively on coding regions and essential splice sites, thereby overlooking other non-coding variants. As a result, intronic variants that can promote mis-splicing events across a range of diseases, including cancer, are yet to be systematically investigated. Such investigations would require both genomic and transcriptomic data, but there currently exist very few datasets that satisfy these requirements. We address this by developing a single-nucleus full-length RNA-sequencing approach that allows for the detection of potentially pathogenic intronic variants. We exemplify the potency of our approach by applying pancreatic cancer tumor and tumor-derived specimens and linking intronic variants to splicing dysregulation. We specifically find that prominent intron retention and pseudo-exon activation events are shared by the tumors and affect genes encoding key transcriptional regulators. Our work paves the way for the assessment and exploitation of intronic mutations as powerful prognostic markers and potential therapeutic targets in cancer.© The Author(s) 2024. Published by Oxford University Press on behalf of NAR Genomics and Bioinformatics.