The role of RNA splicing factors in Ras transformed cells

背景。通过shRNA合成致死筛选，我们已经确定了KRAS突变癌细胞生存所需的一些mRNA剪接因子。目的。在这个项目中，我们的目标是解决以下问题：1)mRNA剪接因子的缺失导致KRAS癌基因合成致死的机制；2)哪些细胞mrna在KRAS突变细胞中被差异剪接；3)与KRAS野生型细胞相比，KRAS突变细胞mRNA剪接模式的变化如何影响其生存能力。重要的材料和方法。1)靶向剪接因子的shrna。2)定量mRNA剪接模式变化的RNA-seq方案。的成就。我们已经确定了一种新的机制，通过这种机制，KRAS基因的剪接是由外显子连接复合物调节的。一份描述我们发现的手稿正在审查中，准备发表。

BACKGROUND. Through a shRNA synthetic lethal screen we have identified a number of mRNA splicing factors that are required for the viability of KRAS mutant cancer cells. PURPOSE. In this project we aim to address the following questions: 1) the mechanism by which depletion of mRNA splicing factors causes synthetic lethality with the KRAS oncogene; 2) which cellular mRNAs are differentially spliced in KRAS mutant cells; 3) how the changes in mRNA splicing pattern in KRAS mutant cells affect their viability compared to KRAS wild type cells. SIGNIFICANT MATERIALS AND METHODS. 1) shRNAs that target splicing factors. 2) RNA-seq protocol for quantifying mRNA splicing pattern changes. ACCOMPLISHMENT. We have identified a novel mechanism by which the splicing of the KRAS gene is regulated by the exon junction complex. A manuscript describing our finding is under review for publication.