title: Novel recombinant hepatitis B virus vectors efficiently deliver protein and RNA encoding genes into primary hepatocytes.
abstract: Hepatitis B virus (HBV) has extremely restricted host and hepatocyte tropism. HBV-based vectors could form the basis of novel therapies for chronic hepatitis B and other liver diseases and would also be invaluable for the study of HBV infection. Previous attempts at developing HBV-based vectors encountered low yields of recombinant viruses and/or lack of sufficient infectivity/cargo gene expression in primary hepatocytes, which hampered follow-up applications. In this work, we constructed a novel vector based on a naturally occurring, highly replicative HBV mutant with a 207-bp deletion in the preS1/polymerase spacer region. By applying a novel insertion strategy that preserves the continuity of the polymerase open reading frame (ORF), recombinant HBV (rHBV) carrying protein or small interfering RNA (siRNA) genes were obtained that replicated and were packaged efficiently in cultured hepatocytes. We demonstrated that rHBV expressing a fluorescent reporter (DsRed) is highly infective in primary tree shrew hepatocytes, and rHBV expressing HBV-targeting siRNA successfully inhibited antigen expression from coinfected wild-type HBV. This novel HBV vector will be a powerful tool for hepatocyte-targeting gene delivery, as well as the study of HBV infection.
authors.affiliation: Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
authors.forename: Ran
authors.initials: R
authors.lastname: Hong
authors.forename: Weiya
authors.initials: W
authors.lastname: Bai
authors.forename: Jianwei
authors.initials: J
authors.lastname: Zhai
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.forename: Xinyan
authors.initials: X
authors.lastname: Li
authors.forename: Jiming
authors.initials: J
authors.lastname: Zhang
authors.forename: Xiaoxian
authors.initials: X
authors.lastname: Cui
authors.forename: Xue
authors.initials: X
authors.lastname: Zhao
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Ye
authors.forename: Qiang
authors.initials: Q
authors.lastname: Deng
authors.forename: Pierre
authors.initials: P
authors.lastname: Tiollais
authors.forename: Yumei
authors.initials: Y
authors.lastname: Wen
authors.forename: Jing
authors.initials: J
authors.lastname: Liu
authors.forename: Youhua
authors.initials: Y
authors.lastname: Xie
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D017353:Gene Deletion; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors; D056945:Hep G2 Cells; D006509:Hepatitis B; D006514:Hepatitis B Surface Antigens / Q000235:genetics / Q000378:metabolism; D006515:Hepatitis B virus / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity*; D022781:Hepatocytes / Q000821:virology*; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D014418:Tupaiidae; D014779:Virus Replication
chemical_list: D006514:Hepatitis B Surface Antigens; D008164:Luminescent Proteins; D011498:Protein Precursors; C413663:fluorescent protein 583; C050806:presurface protein 1, hepatitis B surface antigen
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