用于靶向RNA长扩增子测序(rLAS)的差异剪接工具的计算比较
Computational Comparison of Differential Splicing Tools for Targeted RNA Long-Amplicon Sequencing (rLAS).
作者信息
Ura Hiroki, Hatanaka Hisayo, Togi Sumihito, Niida Yo
机构信息
Center for Clinical Genomics, Kanazawa Medical University Hospital, 1-1 Daigaku, Uchinada, Kahoku 920-0923, Ishikawa, Japan.
Division of Genomic Medicine, Department of Advanced Medicine, Medical Research Institute, Kanazawa Medical University, 1-1 Daigaku, Uchinada, Kahoku 920-0923, Ishikawa, Japan.
出版信息
Int J Mol Sci. 2025 Mar 30;26(7):3220. doi: 10.3390/ijms26073220.
RNA sequencing (RNA-Seq) is a powerful technique for the quantification of transcripts and the analysis of alternative splicing. Previously, our laboratory developed the targeted RNA long-amplicon sequencing (rLAS) method, which has the advantage of allowing deep analysis of targeted specific transcripts. The computational tools for analyzing RNA-Seq data have boosted alternative splicing research by detecting and quantifying splicing events. However, the performance of these splicing tools has not yet been investigated for rLAS. Here, we evaluated the performance of four splicing tools (MAJIQ, rMATS, MISO, and SplAdder) using samples with different types of known splicing events (exon-skipping, multiple-exon-skipping, alternative 5' splicing, and alternative 3' splicing). MAJIQ was able to detect all of the types of events, but it was unable to detect one of the exon-skipping events. On the other hand, rMATS was able to detect all of the exon-skipping events. However, rMATS failed to detect other types of events besides exon-skipping events. Both MISO and SplAdder were unable to detect any of the events. These results indicate that MAJIQ presents better performance for the different types of splicing events in rLAS and that rMATS shows better performance for exon-skipping splicing events.
RNA测序(RNA-Seq)是一种用于转录本定量和可变剪接分析的强大技术。此前,我们实验室开发了靶向RNA长扩增子测序(rLAS)方法,该方法具有能够深入分析靶向特定转录本的优势。用于分析RNA-Seq数据的计算工具通过检测和定量剪接事件推动了可变剪接研究。然而,尚未针对rLAS研究这些剪接工具的性能。在此,我们使用具有不同类型已知剪接事件(外显子跳跃、多外显子跳跃、可变5'剪接和可变3'剪接)的样本评估了四种剪接工具(MAJIQ、rMATS、MISO和SplAdder)的性能。MAJIQ能够检测所有类型的事件,但无法检测到其中一个外显子跳跃事件。另一方面,rMATS能够检测所有外显子跳跃事件。然而,rMATS除了外显子跳跃事件外未能检测到其他类型的事件。MISO和SplAdder均无法检测到任何事件。这些结果表明,MAJIQ在rLAS中对不同类型的剪接事件表现出更好的性能,而rMATS在外显子跳跃剪接事件中表现出更好的性能。