title: CRISPR/Cas9 produces anti-hepatitis B virus effect in hepatoma cells and transgenic mouse.
abstract: Chronic infection of hepatitis B virus (HBV) is at risk of liver cirrhosis and hepatocellular carcinoma and remains one of the major public health problems worldwide. It is a major barrier of persistence HBV cccDNA under current antiviral therapy as novel strategies of disrupting HBV cccDNA is pressing. The (CRISPR)/Cas9 system is presently emerging in gene editing and we also apply it for targeting and deleting the conserved regions of HBV genome. Two homologous sequences of HBV S and X genes were carried with CRISPR/Cas9 endonuclease to build pCas9 constructs, which may mediate anti-HBV effects of in vitro and in vivo systems in this study. The results showed the better anti-HBV productions by pCas9-2 and without significant differences in between Huh7 and HepG2 cells. CRISPR/Cas9 direct cleavage and mutagenesis were further analyzed of in vitro system. In the M-TgHBV mouse model of HBV, injection of pCas9 constructs by hydrodynamics decreased HBsAg of sera and liver HBcAg. In conclusion, this designed CRISPR/Cas9 system can induce anti-HBV effects and potentially consider as a novel therapeutic agent against chronic HBV infection.
authors.affiliation: Laboratory of Molecular Medicine, School of Life Sciences and Technology, Tongji University, Shanghai 200092, China. Electronic address: zhuwei8247@aliyun.com.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhu
authors.affiliation: Laboratory of Molecular Medicine, School of Life Sciences and Technology, Tongji University, Shanghai 200092, China.
authors.forename: Kun
authors.initials: K
authors.lastname: Xie
authors.affiliation: Laboratory of Molecular Medicine, School of Life Sciences and Technology, Tongji University, Shanghai 200092, China.
authors.forename: Yuanjian
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Laboratory of Molecular Medicine, School of Life Sciences and Technology, Tongji University, Shanghai 200092, China.
authors.forename: Le
authors.initials: L
authors.lastname: Wang
authors.affiliation: Laboratory of Molecular Medicine, School of Life Sciences and Technology, Tongji University, Shanghai 200092, China.
authors.forename: Kaiming
authors.initials: K
authors.lastname: Chen
authors.affiliation: Laboratory of Molecular Medicine, School of Life Sciences and Technology, Tongji University, Shanghai 200092, China.
authors.forename: Longzhen
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Laboratory of Molecular Medicine, School of Life Sciences and Technology, Tongji University, Shanghai 200092, China. Electronic address: jfang@Tongji.edu.cn.
authors.forename: Jianmin
authors.initials: J
authors.lastname: Fang
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics / Q000494:pharmacology*; D000076987:CRISPR-Associated Protein 9; D064113:CRISPR-Cas Systems; D045744:Cell Line, Tumor; D004270:DNA, Circular / Q000187:drug effects; D004279:DNA, Viral / Q000187:drug effects; D016503:Drug Delivery Systems; D004720:Endonucleases / Q000235:genetics / Q000494:pharmacology*; D016679:Genome, Viral / Q000187:drug effects; D006509:Hepatitis B / Q000517:prevention & control*; D006515:Hepatitis B virus / Q000187:drug effects / Q000235:genetics*; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic
chemical_list: D000998:Antiviral Agents; D001426:Bacterial Proteins; D004270:DNA, Circular; D004279:DNA, Viral; D000076987:CRISPR-Associated Protein 9; C000606107:Cas9 endonuclease Streptococcus pyogenes; D004720:Endonucleases
keywords: CRISPR/Cas9; HBV; Hydrodynamics; cccDNA