title: Re-Designed Recombinant Hepatitis B Virus Vectors Enable Efficient Delivery of Versatile Cargo Genes to Hepatocytes with Improved Safety.
abstract: Hepatitis B virus (HBV) takes humans as its sole natural host, and productive infection in vivo is restricted exclusively to hepatocytes in the liver. Consequently, HBV-derived viral vectors are attractive candidates for liver-targeting gene therapies. Previously, we developed a novel recombinant HBV vector, designated 5c3c, from a highly replicative clinical isolate. 5c3c was demonstrated to be capable of efficiently delivering protein or RNA expression into infected primary tupaia hepatocytes (PTH), but the design of 5c3c imposes stringent restrictions on inserted sequences, which have limited its wider adoption. In this work, we addressed issues with 5c3c by re-designing the insertion strategy. The resultant vector, designated 5dCG, was more replicative than parental 5c3c, imposed no specific restrictions on inserted sequences, and allowed insertion of a variety of cargo genes without significant loss of replication efficiency. 5dCG-based recombinant HBV effectively delivered protein and RNA expression into infected PTH. Furthermore, due to the loss of functional core ORF, 5dCG vectors depend on co-infecting wild type HBV for replication and efficient expression of cargo genes. Development of the improved 5dCG vector makes wider applications of recombinant HBV possible, while dependence on co-infecting wild type HBV results in improved safety for certain in vivo applications.
authors.affiliation: Key Laboratory of Medical Molecular Virology (MOH &amp; MOE) and Institutes of Biomedical Sciences, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China. bwy.1989@163.com.
authors.forename: Weiya
authors.initials: W
authors.lastname: Bai
authors.affiliation: Key Laboratory of Medical Molecular Virology (MOH &amp; MOE) and Institutes of Biomedical Sciences, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China. cuixiaoxian9@hotmail.com.
authors.forename: Xiaoxian
authors.initials: X
authors.lastname: Cui
authors.affiliation: Key Laboratory of Medical Molecular Virology (MOH &amp; MOE) and Institutes of Biomedical Sciences, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China. chenruidong1000@163.com.
authors.forename: Ruidong
authors.initials: R
authors.lastname: Chen
authors.affiliation: Key Laboratory of Medical Molecular Virology (MOH &amp; MOE) and Institutes of Biomedical Sciences, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China. 13211010047@fudan.edu.cn.
authors.forename: Shuai
authors.initials: S
authors.lastname: Tao
authors.affiliation: Key Laboratory of Medical Molecular Virology (MOH &amp; MOE) and Institutes of Biomedical Sciences, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China. mol725@163.com.
authors.forename: Ran
authors.initials: R
authors.lastname: Hong
authors.affiliation: Department of Infectious Diseases, Huashan Hospital, Fudan University, Shanghai 200040, China. jmzhang@fudan.edu.cn.
authors.forename: Jiming
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Medical Molecular Virology (MOH &amp; MOE) and Institutes of Biomedical Sciences, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China. junqizhang@fudan.edu.cn.
authors.forename: Junqi
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Medical Molecular Virology (MOH &amp; MOE) and Institutes of Biomedical Sciences, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China. yxwang712@yahoo.com.
authors.forename: Yongxiang
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Key Laboratory of Medical Molecular Virology (MOH &amp; MOE) and Institutes of Biomedical Sciences, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China. yhxie@fudan.edu.cn.
authors.forename: Youhua
authors.initials: Y
authors.lastname: Xie
authors.affiliation: Key Laboratory of Medical Molecular Virology (MOH &amp; MOE) and Institutes of Biomedical Sciences, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China. liujing212@fudan.edu.cn.
authors.forename: Jing
authors.initials: J
authors.lastname: Liu
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D005822:Genetic Vectors; D006515:Hepatitis B virus / Q000235:genetics*; D022781:Hepatocytes / Q000821:virology*; D014161:Transduction, Genetic; D014417:Tupaia
keywords: HBV; gene therapy; hepatocyte; viral vector