基于CRISPR文库筛选发现SNRPC基因通过RNA可变剪接调控三阴性乳腺癌发展的分子机制研究

三阴性乳腺癌(TNBC)因自身病因的复杂性且缺乏明确治疗靶点，成为当今乳腺癌研究的热点之一。已有的乳腺癌病因学研究大多从基因组或转录组的角度展开，而从RNA可变剪接层面开展的探索研究较为匮乏。本项目基于申请者对RNA可变剪切的工作积累，利用针对RNA结合蛋白的CRISPR文库进行高通量筛选，发现了小核糖核蛋白SNRPC对于三阴性乳腺癌的进展具有重要调控功能。在此基础上，本项目拟采用多种先进的实验技术，系统性地解析SNRPC通过RNA可变剪切调控三阴性乳腺癌进展的分子机制，包括：1)明确SNRPC通过U1剪接体对RNA可变剪接的调控作用；2)阐明SNRPC通过RNA可变剪接影响三阴性乳腺癌生存信号的分子机制；3)探索基因组和表观基因组信号共同调控RNA剪接体染色质定位的分子机制。本课题将揭示RNA可变剪接在调控三阴性乳腺癌进展中的重要性，为发现三阴性乳腺癌的新型靶标奠定理论和实验基础。

Triple-negative breast cancer (TNBC) has become one of the major challenges in breast cancer research, because of its complexity of etiology and lack of treatment targets. Most of the existing studies on tumor etiology are conducted at the genomic or transcriptional levels, while the research on RNA splicing is relatively scarce. In this project, we found that SNRPC was an important RNA splicing regulator and controled the development of triple negative breast cancer, via using high-throughput CRISPR screening targeting RNA binding proteins (RBPs). On this basis, the project plans to systematically analyze the molecular mechanism of SNRPC in regulating the progress of TNBC through RNA alternative splicing, including: 1) Clarifying the regulatory effect of SNRPC on RNA alternative splicing through U1 spliceosome; 2) clarifying the molecular mechanism of SNRPC affecting tumor survival signal; 3) exploring the roles of genomic and epigenetic signaling in regulating the chromatin localization of spliceosomes. This study revealed the importance of RNA alternative splicing in the regulation of TNBC development, and laid an experimental and theoretical foundation to discover the new targets in TNBCs.