title: An efficient antiviral strategy for targeting hepatitis B virus genome using transcription activator-like effector nucleases.
abstract: The hepatitis B virus (HBV) is a DNA virus that can cause chronic hepatitis B (CHB) in humans. Current therapies for CHB infection are limited in efficacy and do not target the pre-existing viral genomic DNA, which are present in the nucleus as a covalently closed circular DNA (cccDNA) form. The transcription activator-like (TAL) effector nucleases (TALENs) are newly developed enzymes that can cleave sequence-specific DNA targets. Here, TALENs targeting the conserved regions of the viral genomic DNA among different HBV genotypes were constructed. The expression of TALENs in Huh7 cells transfected with monomeric linear full-length HBV DNA significantly reduced the viral production of HBeAg, HBsAg, HBcAg, and pgRNA, resulted in a decreased cccDNA level and misrepaired cccDNAs without apparent cytotoxic effects. The anti-HBV effect of TALENs was further demonstrated in a hydrodynamic injection-based mouse model. In addition, an enhanced antiviral effect with combinations of TALENs and interferon-α (IFN-α) treatment was observed and expression of TALENs restored HBV suppressed IFN-stimulated response element-directed transcription. Taken together, these data indicate that TALENs can specifically target and successfully inactivate the HBV genome and are potently synergistic with IFN-α, thus providing a potential therapeutic strategy for treating CHB infection.
authors.affiliation: Key Laboratory of Medical Molecular Virology, School of Basic Medical Sciences, Shanghai Medical College of Fudan University, Shanghai, China; Shanghai Public Health Clinical Center, Fudan University, Shanghai, China.
authors.forename: Jieliang
authors.initials: J
authors.lastname: Chen
authors.affiliation: Key Laboratory of Medical Molecular Virology, School of Basic Medical Sciences, Shanghai Medical College of Fudan University, Shanghai, China; Shanghai Public Health Clinical Center, Fudan University, Shanghai, China.
authors.forename: Wen
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Medical Molecular Virology, School of Basic Medical Sciences, Shanghai Medical College of Fudan University, Shanghai, China.
authors.forename: Junyu
authors.initials: J
authors.lastname: Lin
authors.affiliation: Key Laboratory of Medical Molecular Virology, School of Basic Medical Sciences, Shanghai Medical College of Fudan University, Shanghai, China; Shanghai Public Health Clinical Center, Fudan University, Shanghai, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Wang
authors.affiliation: Shanghai Public Health Clinical Center, Fudan University, Shanghai, China.
authors.forename: Min
authors.initials: M
authors.lastname: Wu
authors.affiliation: Key Laboratory of Medical Molecular Virology, School of Basic Medical Sciences, Shanghai Medical College of Fudan University, Shanghai, China; Institutes of Medical Microbiology and Biomedical Sciences, Fudan University, Shanghai, China.
authors.forename: Cuncun
authors.initials: C
authors.lastname: Chen
authors.affiliation: Shanghai Public Health Clinical Center, Fudan University, Shanghai, China.
authors.forename: Ye
authors.initials: Y
authors.lastname: Zheng
authors.affiliation: Shanghai Public Health Clinical Center, Fudan University, Shanghai, China.
authors.forename: Xiuhua
authors.initials: X
authors.lastname: Peng
authors.affiliation: Key Laboratory of Medical Molecular Virology, School of Basic Medical Sciences, Shanghai Medical College of Fudan University, Shanghai, China.
authors.forename: Jianhua
authors.initials: J
authors.lastname: Li
authors.affiliation: Key Laboratory of Medical Molecular Virology, School of Basic Medical Sciences, Shanghai Medical College of Fudan University, Shanghai, China; Shanghai Public Health Clinical Center, Fudan University, Shanghai, China; Institutes of Medical Microbiology and Biomedical Sciences, Fudan University, Shanghai, China. Electronic address: zhyuan@shaphc.org.
authors.forename: Zhenghong
authors.initials: Z
authors.lastname: Yuan
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000008:administration & dosage; D002460:Cell Line; D004270:DNA, Circular / Q000378:metabolism; D003851:Deoxyribonucleases / Q000235:genetics / Q000378:metabolism*; D004195:Disease Models, Animal; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D016679:Genome, Viral; D006515:Hepatitis B virus / Q000235:genetics*; D019694:Hepatitis B, Chronic / Q000378:metabolism* / Q000628:therapy / Q000821:virology*; D006801:Humans; D016898:Interferon-alpha / Q000008:administration & dosage; D051379:Mice; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D011485:Protein Binding; D014764:Viral Proteins / Q000378:metabolism
chemical_list: D000998:Antiviral Agents; D004270:DNA, Circular; D016898:Interferon-alpha; D014764:Viral Proteins; D003851:Deoxyribonucleases
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