前列腺癌（Prostate Cancer,PCa）发生发展中伴随大量的基因突变和融合基因事件，但其中关键的功能性变异及相关作用机制并不完全清楚。建立针对高频突变基因的高通量功能性筛选方案，对于发现及研究新的关键作用基因可能具有重要价值。本课题预实验将大样本高通量测序与CRISPR-Cas9筛选策略结合，着眼于染色体易位和高频突变基因，构建小型sgRNA文库，鉴定到可能在PCa中起重要作用的新候选基因UBXN4。敲除UBXN4后的PCa细胞增殖加快、成瘤能力增加。敲除细胞测序后发现差异基因明显富集到MAPK通路，结合生信分析结果我们初步提出UBXN4通过HRAS-MAPK通路影响PCa进展的科学假设。本项目将进一步通过CRISPR-Cas9，CoIP，蛋白质谱等技术在多个细胞系，以及动物和临床样本水平证实该假设。拟为深化对PCa的认识，促进未来PCa诊疗发展提供实验参考。

Prostate Cancer (PCa) is accompanied by a large number of gene mutations and fusion gene events, but the key functional variations and related mechanisms are not fully clear. Establishing a high-throughput functional screening scheme for high-frequency mutation genes may be of great value for discovering and studying new critical genes.In our preliminary experiment, we combined the second generation sequencing technology with the screening strategy of CRISPR gene library. On the basis of large sample high throughput sequencing, we focused on chromosome translocation and high frequency mutation genes, constructed a small sgRNA library, and screened genes that play roles inPCa. And we identified UBXN4 may play an important role in PCa. After knocking out UBXN4, PCa cells proliferated faster and their tumorigenicity increased. High throughput sequencing results shown that the differentially expressed genes after UBXN4 knock-out were obviously enriched in the MAPK pathway. Combined with the results of bioinformatics analysis and experimental result, we preliminarily proposed the scientific hypothesis that UBXN4 affected the progression of cancer through HRAS-MAPK pathway. This project will further validate this hypothesis through CRISPR-Cas9, CoIP, and protein profiling techniques in multiple cell lines, as well as in animal and clinical samples. It is intended to provide experimental reference for deepening the understanding of PCa and promoting the development of UBXN4-related diagnosis and treatment of PCa in the future.