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Development of hybrid baculovirus vectors for artificial MicroRNA delivery and prolonged gene suppression

Authors

  • Chiu-Ling Chen,

    1. Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan, 300; telephone: (886)3-5718245; fax: (886)3-5715408;
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  • Wen-Yi Luo,

    1. Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan, 300; telephone: (886)3-5718245; fax: (886)3-5715408;
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  • Wen-Hsin Lo,

    1. Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan, 300; telephone: (886)3-5718245; fax: (886)3-5715408;
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  • Kun-Ju Lin,

    1. Department of Nuclear Medicine and Molecular Imaging Center, Chang Gung Memorial Hospital, Taoyuan, Taiwan
    2. Department of Medical Imaging and Radiological Sciences, Chang Gung University, Taoyuan, Taiwan
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  • Li-Yu Sung,

    1. Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan, 300; telephone: (886)3-5718245; fax: (886)3-5715408;
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  • Yung-Shen Shih,

    1. Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan, 300; telephone: (886)3-5718245; fax: (886)3-5715408;
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  • Yu-Han Chang,

    Corresponding author
    1. College of Medicine, Chang Gung University, Taoyuan, Taiwan, 333; telephone: (886)3-328-1200; fax: (886)3-3278113;
    2. Department of Orthopaedic, Chang Gung Memorial Hospital, Taoyuan, Taiwan
    • College of Medicine, Chang Gung University, Taoyuan, Taiwan, 333; telephone: (886)3-328-1200; fax: (886)3-3278113;.
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  • Yu-Chen Hu

    Corresponding author
    1. Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan, 300; telephone: (886)3-5718245; fax: (886)3-5715408;
    • Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan, 300; telephone: (886)3-5718245; fax: (886)3-5715408;.
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  • Conflict of interest: None.

Abstract

MicroRNA (miRNA) plays essential roles in regulating gene expression, but miRNA delivery remains a hurdle, thus entailing a vector system for efficient transfer. Baculovirus emerges as a promising gene delivery vector but its inherent transient expression restricts its applications in some scenarios. Therefore, this study primarily aimed to develop baculovirus as a miRNA expression vector for prolonged gene suppression. We constructed recombinant baculoviruses carrying artificial egfp-targeting miRNA sequences within the miR155 backbone, which after expression by the cytomegalovirus promoter could knockdown the enhanced green fluorescent protein (EGFP) expression in a sequence- and dose-dependent manner. By swapping the mature miRNA sequences, the baculovirus miRNA shuttle effectively repressed the overexpression of endogenous TNF-α in arthritic synoviocytes without inducing apoptosis. To prolong the baculovirus-mediated expression, we further developed a hybrid baculovirus vector that exploited the Sleeping Beauty (SB) transposon for gene integration and sustained miRNA expression. The hybrid baculovirus vector that combined the miR155 scaffold and SB transposon effectively repressed the transgene expression for a prolonged period of time, hence diversifying the applications of baculovirus to indications necessitating prolonged gene regulation such as arthritis. Biotechnol. Bioeng. 2011;108: 2958–2967. © 2011 Wiley Periodicals, Inc.

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