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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.<\/jats:p>","DOI":"10.1128\/jvi.03328-12","type":"journal-article","created":{"date-parts":[[2013,4,4]],"date-time":"2013-04-04T06:59:59Z","timestamp":1365058799000},"page":"6615-6624","update-policy":"http:\/\/dx.doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":40,"title":["Novel Recombinant Hepatitis B Virus Vectors Efficiently Deliver Protein and RNA Encoding Genes into Primary Hepatocytes"],"prefix":"10.1128","volume":"87","author":[{"given":"Ran","family":"Hong","sequence":"first","affiliation":[{"name":"Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China"},{"name":"Key Laboratory of Medical Molecular Virology (Ministry of Health and Ministry of Education) and Institutes of Biomedical Sciences, Shanghai Medical College, Fudan University, 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