SWAN Institute of Biomedical and Life Sciences, School of Biological Sciences, Royal Holloway, University of London, Egham, Surrey, UK.
In Vivo. 2009 Nov-Dec;23(6):885-93.
To improve liver-directed retroviral-mediated gene transfer, we injected C57/BL10 mice intravenously with three adenoviral vectors encoding retroviral vector genome and structural components: AdGagPol expressing the respective structural genes of Moloney murine leukaemia virus, Ad10A1Env expressing the 10A1 envelope protein of 10A1-MuLV, and AdLEIN, encoding the LEIN retrovirus genome, expressing green fluorescence protein (eGFP) and the neomycin resistance gene.
The extent of eGFP expression was determined after 1 and 15 weeks by fluorescence microscopy and FACS analysis. Proviral integration was determined by a novel PCR-based technique.
Hepatocytes infected with all three Ad vectors generated LEIN retrovirus after one week and in situ transduction of neighbouring cells resulted in stable proviral integration associated with eGFP expression ranging from 4.3% to 20.5% in different liver cell populations 15 weeks post-infection.
Hybrid adeno-retroviral vectors can be efficiently used to improve the efficiency of retroviral-mediated gene transfer to the liver.
为了提高肝靶向逆转录病毒介导的基因转移，我们将三种腺病毒载体（分别编码逆转录病毒载体基因组和结构成分）静脉注射到 C57/BL10 小鼠体内：AdGagPol 表达 Moloney 鼠白血病病毒的相应结构基因，Ad10A1Env 表达 10A1-MuLV 的 10A1 包膜蛋白，以及 AdLEIN，编码 LEIN 逆转录病毒基因组，表达绿色荧光蛋白（eGFP）和新霉素抗性基因。
通过荧光显微镜和 FACS 分析，在第 1 周和第 15 周时确定 eGFP 的表达程度。通过一种新的基于 PCR 的技术确定前病毒整合。
感染所有三种 Ad 载体的肝细胞在一周后产生 LEIN 逆转录病毒，并且原位转导邻近细胞导致稳定的前病毒整合，与感染后 15 周不同肝实质细胞群中 eGFP 表达范围为 4.3%至 20.5%相关。
杂交腺病毒 - 逆转录病毒载体可有效地用于提高逆转录病毒介导的基因转移到肝脏的效率。
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