伴随RNA 干扰、CRISPR基因编辑等生物技术日新月异的发展，使得从根本上清除寄存在人体的乙肝病毒（Hepatitis B Virus, HBV）成为可能，而如何将这些治疗基因靶向输送到肝细胞内并高效表达成为能否治愈乙肝的关键。基于前期研究建立的腺相关病毒（adeno-associated virus, AAV）的非天然氨基酸定点标记新技术，本课题拟进一步研究揭示病毒载体修饰后的衣壳结构与感染活性及靶向性的构效关系，发现病毒载体的可靶向修饰位点，进而通过定点共价偶联N-乙酰半乳糖构建肝靶向AAV载体，将由肝特异启动子引导的CRISPR/Cas9基因和高效抑制HBV复制的microRNA基因优化组合，靶向转运至肝细胞内长期表达。利用基因编辑和RNA干扰的双重效果，从根本上抑制并清除寄存在人体的HBV。该课题的成功实施，将造福亿万乙肝患者，也会使基因治疗向更高效、安全的方向迈出关键一步。

Along with the rapid development of RNA interference, CRISPR gene editing and other biological technology, thoroughly clearance of the hepatitis B virus stored in human is possible, and how these genes are selectively delivered to the liver cells and high expression become the key of hepatitis B treatment. Based on the new technology of unnatural amino acid site-specific modification of adeno-associated virus, this study intends to further study the structure-activity relationship between the modification of viral capsid structure and the active infection and targeting of viral vector and find the proper sites for viral targeting modification. Through site directed covalent coupling of N-acetylgalactosamine to adeno-associated virus, the liver targeting vector is constructed. Using this vector, the liver specific promoter guided CRISPR / cas9 gene and microRNA gene of HBV replication inhibition are transferred to the liver cells for long-term expression. Then, we can fundamentally remove the HBV stored in human by the double effect of CRISPR gene editing and RNA interference suppression. When this study has been successfully completed, it will not be only benefit for the millions of patients with hepatitis B, also makes gene therapy more efficiency and safe.