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Water-Soluble and Low Molecular Weight Chitosan-Based Plasmid DNA Delivery

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Abstract

Purpose. Chitosan, a natural cationic polysaccharide, is a candidate non-viral vector for gene delivery because of its high positive charges and low cytotoxicity. In this study, low molecular weight chitosan (LMWC, molecular weight of 22 kDa) was characterized and evaluated as a gene carrier.

Methods. Plasmid/LMWC complex was analyzed in 1% agarose gel electrophoresis. To confirm that the LMWC protected plasmids from nuclease, DNase I protection assays were performed. pSV-β-galactosidase plasmid/LMWC complex was transfected into 293T cells and transfection efficiency was evaluated by β-galactosidase assay. Cytotoxicity of LMWC was determined by MTT assay.

Results. Unlike high molecular weight chitosan (HMWC), LMWC is highly water soluble, and can form complex with plasmids in physiological buffer. The plasmid DNA was completely retarded at a weight ratio of 1:2 (plasmid:LMWC) in 1% agarose gel. DNase I protection assay showed that plasmids were protected from DNase I over 60 min. The most efficient transfection was obtained at a weight ratio of 1:3 (plasmid:LMWC). The transfection efficiency of LMWC was significantly higher than naked DNA and higher than poly-L-lysine (PLL). MTT assay showed that LMWC was less cytotoxic than PLL.

Conclusions. LMWC is non-toxic and has higher transfection efficiency than PLL. Therefore, LMWC will be useful in the development of safe gene carriers.

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Authors and Affiliations

  1. Department of Pharmaceutics and Pharmaceutical Chemistry, Center for Controlled Chemical Delivery, University of Utah, 30 S 2000 E RM 201, Salt Lake City, Utah, 84112-5820

    Minhyung Lee, Youngmin Kwon, Jae Joon Koh & Sung Wan Kim

  2. Department of Polymer Science and Engineering, Sunchon National University, Chonnam, 540-742, Korea

    Jae-Woon Nah

  3. Department of Internal Medicine, College of Medicine, Inje University, Seoul, 130-707, Korea

    Kyung Soo Ko

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  1. Minhyung Lee
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Lee, M., Nah, JW., Kwon, Y. et al. Water-Soluble and Low Molecular Weight Chitosan-Based Plasmid DNA Delivery. Pharm Res 18, 427–431 (2001). https://doi.org/10.1023/A:1011037807261

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