Liu Xing, Hao Ruidong, Chen Shuliang, Guo Deyin, Chen Yu
State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, Hubei, 430072, PR China.
School of Basic Medical Sciences, Wuhan University, Wuhan, Hubei, 430072, PR China.
J Gen Virol. 2015 Aug;96(8):2252-2261. doi: 10.1099/vir.0.000159. Epub 2015 Apr 22.
Hepatitis B virus (HBV) remains a global health threat as chronic HBV infection may lead to liver cirrhosis or cancer. Current antiviral therapies with nucleoside analogues can inhibit the replication of HBV, but do not disrupt the already existing HBV covalently closed circular DNA. The newly developed CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated 9) system is a powerful tool to target cellular genome DNA for gene editing. In order to investigate the possibility of using the CRISPR/Cas9 system to disrupt the HBV DNA templates, we designed eight guide RNAs (gRNAs) that targeted the conserved regions of different HBV genotypes, which could significantly inhibit HBV replication both in vitro and in vivo. Moreover, the HBV-specific gRNA/Cas9 system could inhibit the replication of HBV of different genotypes in cells, and the viral DNA was significantly reduced by a single gRNA/Cas9 system and cleared by a combination of different gRNA/Cas9 systems.
乙型肝炎病毒（HBV）仍然是一个全球性的健康威胁，因为慢性HBV感染可能导致肝硬化或癌症。目前使用核苷类似物的抗病毒疗法可以抑制HBV的复制，但不会破坏已存在的HBV共价闭合环状DNA。新开发的CRISPR（成簇规律间隔短回文重复序列）/Cas9（CRISPR相关蛋白9）系统是一种用于靶向细胞基因组DNA进行基因编辑的强大工具。为了研究使用CRISPR/Cas9系统破坏HBV DNA模板的可能性，我们设计了8种靶向不同HBV基因型保守区域的引导RNA（gRNA），它们在体外和体内均可显著抑制HBV复制。此外，HBV特异性gRNA/Cas9系统可抑制细胞中不同基因型HBV的复制，单个gRNA/Cas9系统可显著减少病毒DNA，不同gRNA/Cas9系统联合使用可清除病毒DNA。
