重组腺相关病毒介导nSpRY-ABEmax系统靶向修复Leptin基因对ob/ob肥胖小鼠的治疗研究

基于CRISPR/Cas系统相关基因编辑技术的迅速发展为研究疾病提供了重要思路，已在多种遗传性疾病小鼠模型中实现了成体治疗。肥胖症是由遗传、环境和行为相互作用的结果，早发的重度肥胖主要是由基因突变导致的遗传性肥胖。目前，利用单碱基编辑技术成体治疗遗传性肥胖的研究还未见报道。ob/ob小鼠是其Leptin基因发生C到T突变产生终止密码子，从而导致肥胖。本研究拟以ob/ob小鼠模型为研究对象，通过尾静脉注射双rAAV介导nSpRY-ABEmax和Leptin sgRNA，对小鼠脂肪细胞Leptin基因进行修复，从而治疗肥胖症；并通过扩增子、转录组和全基因组测序分析，评估nSpRY-ABEmax在体外和体内的脱靶效应。本研究为基因突变导致的遗传性肥胖提供一种新的治疗策略，同时对基于SpRY的碱基编辑器的临床转化应用提供重要理论基础。

The rapid development of CRISPR/Cas-based gene editing technology has provided important ideas for the study of diseases, and it has been applied in the treatment of genetic diseases in adult mouse models. Obesity is the result of interaction of genetics, environment and behavior. The early onset of severe obesity is mainly inherited obesity caused by genetic mutations. So far, the use of base editing for treatment of genetically obese mice has not yet been reported. The obesity occurred in ob/ob mice is due to C to T mutations of Leptin gene to produce a stop codon. In this study, we will take the ob/ob mouse model as research object, deliver dual-rAAV-mediated nSpRY-ABEmax and Leptin sgRNA into adipocyte to correct Leptin gene by tail-vein injection, and explore the feasibility of gene editing in the treatment of inherited obesity. And we will evaluate off-target effects by amplicon, transcriptome and whole genome sequencing analysis. This study will provide a new treatment strategy for genetic obesity caused by gene mutations and provide an important theoretical basis for the clinical translational application of SpRY-based base editing.