新型CRISPR筛选平台的搭建及其在基因功能与染色质调控研究中的应用

基因组计划完成后，系统性的阐明基因在生理、病理过程中的功能成为生物学研究的重要问题。近年来兴起的CRISPR筛选技术能够从全基因组范围内对基因进行敲除或者激活，从而为这一科学问题提供了强大的手段。目前的CRISPR筛选技术由于其工作原理的限制，还需要进一步提高准确率，提高其适用范围。鉴此，我们拟在前期成功搭建于高通量基因筛选平台（Nature 2014，Nat Biotechnol 2016, Nat Biotechnol 2018）的基础上，结合生物信息学手段，建立一整套适用于染色质-基因-基因关键位点的新型高通量功能性筛选技术，并应用于鉴定关键染色体结构调控序列、发现细胞生存和增殖关键基因、构建药物靶点基因的高分辨率功能性图谱等研究，以期从系统水平发现特定条件下的基因及调控元件的功能，并为相关研究提供技术平台。

Beyond genome sequencing, systematically decoding gene function in a specific physiological or pathological process is one of the overarching themes in biological research. Recently developed CRISPR screen enables genome-wide knockout or activation of genes, offering unprecedented opportunities for functional genomics studies. However, current CRISPR screen falls short of precision and sensitivity due to the nature of CRISPR knockout that may give rise to cellular toxicity. Moreover, CRISPR screen is laborious and largely confined to coding genes. In the current research, we propose to develop novel CRISPR screen strategies that are capable of decoding gene functions and functional elements, even in a single-base resolution. By harnessing these methodologies, we set out to identify functional elements that modulate chromosome structure, interrogate genes that are important for cell survival or proliferation, and fine-map drug target genes. As such, we will obtain functional big data for deciphering gene function and provide novel platforms for functional genomics.