低脱靶Cas12a基因编辑系统的分子设计

CRISPR/Cas技术已经在生物医学研究中得到了广泛的应用。多项研究证实，CRISPR/Cas9和CRISPR/Cas12a编辑系统都存在脱靶效应，极大增加了在人体疾病治疗中的风险。进一步研究表明，Cas12a编辑系统相比Cas9编辑系统具有更高的保真性。本研究将Cas12a作为研究对象，在鉴定新的xCas12a的基础上采用分子生物学，细胞生物学，结构生物学，高通量测序，生物信息学等科学方法探究其PAM识别特征，基因编辑功能，全基因组脱靶效应，并基于结构设计低脱靶的xCas12a编辑系统。此项研究将为了解基因编辑系统脱靶效应的分子机制提供重要帮助，同时也为基因编辑提供了更多的备选工具，推动基因编辑技术在疾病治疗中的应用。

CRISPR/Cas technology has been widely used in biomedical research. Many studies have confirmed that both CRISPR/Cas9 and CRISPR/Cas12a editing systems have off-target effects, which greatly increase the risk in the treatment of human diseases. Further studies showed that the Cas12a editing system has higher fidelity than the Cas9 editing system. This study will focus on CRISPR/Cas12a, explore its PAM recognition characteristics and gene editing functions on the basis of the newly identified xCas12a, and design high-fidelity variants based on the structure. The study will provide important help in understanding the molecular mechanism of off-target effects of gene editing systems, and will also provide more alternative tools for gene editing, which may promote the application of gene editing technology in disease treatment.