2015年4月3日，我所熊思东教授在Antiviral Research杂志发表了题为“Targeting hepatitis Bvirus cccDNA by CRISPR/Cas9 nuclease efficiently inhibits viral replication” 的研究论文，报道了一种治疗慢性HBV感染的新策略。
董春升副教授和本科生璩良为该论文的共同第一作者。参与该论文研究的还有卫林老师和董元舒副教授等。熊思东教授和董春升副教授为该论文的通讯作者。该项研究由国家重点基础研究发展计划(2013CB531502，2013CB530501)，国家自然科学基金(31270977，81072413，81101257，31170878)，国家教育部留学回国人员科研启动金（董春升），江苏省“攀登”项目(BK2010004)，江苏省“333高层次人才培养工程”，江苏省高校青蓝工程，江苏省高校优势学科建设工程(PAPD)，江苏省大学生创新训练项目(201310285046Z) 等基金资助。
文章摘要：
Chronic hepatitis B virus (HBV) infection causes liver cirrhosis andhepatocellular carcinoma and remains a serious health problem worldwide.Covalently closed circular DNA (cccDNA) in the liver cell nucleus sustains HBVinfection. Major treatments for HBV infection include the use of interferon-aand nucleotide analogs, but they cannot eradicate cccDNA. As a novel tool forgenome editing, clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associatedprotein 9 (Cas9) system developed from bacteria can be used to accurately andefficiently engineer and modify genomic DNA. In this study, the CRISPR/Cas9system was used to target the HBV genome and efficiently inhibit HBV infection.We synthesized four single-guide RNAs (sgRNAs) targeting the conserved regionsof HBV. The expression of these sgRNAS with Cas9 reduced the viral productionin Huh7 cells as well as in HBV-replication cell HepG2.2.15. We furtherdemonstrated that CRISPR/Cas9 direct cleavage and cleavage-mediated mutagenesisoccurred in HBV cccDNA of transfected cells. In the new mouse model carryingHBV cccDNA, injection of sgRNA–Cas9 plasmids via rapid tail vein resulted in the low level of cccDNA and HBVprotein. In conclusion, the designed CRISPR/Cas9 system can accurately andefficiently target HBV cccDNA and inhibit HBV replication. This system may beused as a novel therapeutic strategy against chronic HBV infection.