Abstract:
Alternative splicing (AS) is an important mechanism contributing to stress-induced regulation of gene expression and proteome diversity. Massive sequencing technologies allow the identification of transcripts generated via stress-responsive AS, potentially important for adaptation to stress conditions. Several bioinformatics tools have been developed to identify differentially expressed alternative splicing events/transcripts from RNA-sequencing results. This chapter describes a detailed protocol for differential alternative splicing analysis using the rMATS tool. In addition, we provide guidelines for validation of the detected splice variants by qRT-PCR based on the obtained output files.
摘要:
选择性剪接(AS)是促进应激诱导的基因表达和蛋白质组多样性调节的重要机制。大规模测序技术允许鉴定通过应激反应性AS产生的转录本,对适应压力条件可能很重要。已经开发了几种生物信息学工具来从RNA测序结果中鉴定差异表达的可变剪接事件/转录物。本章介绍了使用rMATS工具进行差分选择性剪接分析的详细协议。此外,我们根据获得的输出文件提供了通过qRT-PCR验证检测到的剪接变体的指南.