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  • Title: Engineered long terminal repeats of retroviral vectors enhance transgene expression in hepatocytes in vitro and in vivo.
    Author: Yamaguchi K, Itoh K, Ohnishi N, Itoh Y, Baum C, Tsuji T, Nagao T, Higashitsuji H, Okanoue T, Fujita J.
    Journal: Mol Ther; 2003 Nov; 8(5):796-803. PubMed ID: 14599813.
    Abstract:
    To analyze the important elements for retroviral expression in hepatocytes, cis-acting elements in the U3 region of the long terminal repeat (LTR) of the polycythemic strain of spleen focus-forming virus (SFFVp) were analyzed in a hepatocellular carcinoma cell line. Two cis-acting elements located within the upstream region of the direct repeat, which positively regulated retroviral expression, were identified. Transcription factors NFAT5 and Sp1, which are ubiquitously expressed in a variety of tissues, bound to these elements. To increase specificity without lowering the potency of retroviral expression in hepatocytes, these elements were replaced by a sequence derived from the hepatitis B virus enhancer II region. Novel vectors, SF-Hep3 and SF-Hep5 (SFFVp-based vector for hepatocytes 3 and 5), were developed with these engineered LTRs. The engineered LTRs of these vectors enhanced the retroviral expression only in hepatocellular carcinoma cell lines in vitro. These vectors also increased transgene expression 4- to 9-fold or 3.5- to 5-fold in comparison with a Moloney murine leukemia virus-based vector or a vector containing the wild-type LTR of SFFVp, respectively, in murine hepatocytes in vivo.
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