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Amphiphilic peptides with arginines and valines for the delivery of plasmid DNA

Authors

  • Dong-Wook Ryu,

    1. Department of Bioengineering, College of Engineering, Hanyang University, Seoul 133-791, Korea
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  • Hyun Ah Kim,

    1. Department of Bioengineering, College of Engineering, Hanyang University, Seoul 133-791, Korea
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  • Hojung Song,

    1. Department of Bioengineering, College of Engineering, Hanyang University, Seoul 133-791, Korea
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  • Soonhag Kim,

    1. Laboratory of Molecular Imaging, Department of Applied Bioscience, CHA Stem Cell Institute, CHA University, Seoul 135-081, Korea
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  • Minhyung Lee

    Corresponding author
    1. Department of Bioengineering, College of Engineering, Hanyang University, Seoul 133-791, Korea
    2. Institute for Bioengineering and Biopharmaceutical Research, Hanyang University, Seoul 133-791, Korea
    3. Institute of Aging Society, Hanyang University, Seoul 133-791, Korea
    • Department of Bioengineering, College of Engineering, Hanyang University, 17 Haengdang-dong, Seongdong-gu, Seoul 133-791, Korea.
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Abstract

A non-toxic and efficient gene carrier is one requirement for clinical gene therapy. In this study, amphiphilic peptides composed of arginines and valines were synthesized and characterized as plasmid DNA (pDNA) carriers. The peptides have a cationic region containing 1–4 arginines and a hydrophobic region containing 6 valines. The arginine–valine peptides (RV peptides) formed micelles in aqueous solution with a critical micelle concentration (CMC) of 1.35 mg/ml. In gel retardation assay, the RV peptides retarded all pDNA at weight ratios (pDNA:RV peptide) of 1:3 for R1V6, 1:2 for R2V6 and R3V6, and 1:1 for R4V6. A heparin competition assay showed that the R3V6 peptide formed tighter complexes with pDNA than poly-L-lysine (PLL). In vitro transfection assay into HEK293 cells showed that the R1V6 and R2V6 peptides had the highest transfection efficiencies at 1:30 weight ratios (pDNA:RV peptide), while the R3V6 and R4V6 peptides had the highest efficiencies at 1:20 weight ratios. Under optimal conditions, the R3V6 peptide had the highest transfection efficiency of all the RV peptides and PLL. MTT assay showed that the RV peptides did not have any detectable toxicity to cells. Therefore, the RV peptide may be useful for the development of non-toxic gene carriers. J. Cell. Biochem. 112: 1458–1466, 2011. © 2011 Wiley-Liss, Inc.

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