Construction and characterization of liposomal magnetofection system in pig kidney cells
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Genetically Modified Organisms Breeding Major Projects (No. 2009ZX08010-006B), Basic Scientific Research Fund of National Nonprofit Institutes (No. BSRF 201108).

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    Abstract:

    Magnetic nano gene vector is one of the non-viral gene vectors, modified by functional group to bind cationic transfect reagents. Coupling magnetofection with the universal lipofection we developed a novel somatic cell transfection method as the so-called liposomal magnetofection (LMF). This approach is potential to provide somatic cell cloning with stable genetic cell lines to cultivate transgenic animals. In order to construct such liposomal magnetic gene vectors complexes system, we used nano magnetic gene vector to combine with liposomal cationic transfect reagents by molecular self-assembly. This vectors system successfully carried exogenous gene and then transfected animal somatic cells. Here, we conducted atomic force microscopy (AFM), zeta potential-diameter analysis and other characterization experiments to investegate the size distribution and morphology of magnetic nanoparticles, the way of the vectors to load and concentrate DNA molecules. Our data reveal that, the LMF of Pig Kidney cells exhibited higher transfection efficiency comparing with the transfection mediated by the commercial lipofectamine2000. Moreover, LMF method overcomes the constraint of transient expression mediated by lipofection. Meanwhile, MTT assay showed low cytotoxicity of LMF. Hence, LMF is a feasible, low cytotoxic and effective method of cell transfection.

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陈文杰,崔海信,赵翔,崔金辉,王琰,孙长娇. 猪肾细胞脂质磁转染系统的构建与表征[J]. Chinese Journal of Biotechnology, 2014, 30(6): 972-981

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History
  • Received:September 13,2013
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  • Online: June 06,2014
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