Enhancement of Gene Delivery Using Novel Homodimeric Tat Peptide Formed by Disulfide Bond

Title
Enhancement of Gene Delivery Using Novel Homodimeric Tat Peptide Formed by Disulfide Bond
Author(s)
이수진[이수진]윤성화[윤성화]도경오
Keywords
HUMAN IMMUNODEFICIENCY VIRUS; CATIONIC LIPOSOMES; CELLULAR UPTAKE; PLASMID DNA; PROTEIN; TRANSFECTION; COMPLEXES; CARRIERS; THERAPY; CANCER
Issue Date
201108
Publisher
KOREAN SOC MICROBIOLOGY & BIOTECHNOLOGY
Citation
JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY, v.21, no.8, pp.802 - 807
Abstract
Cationic liposomes have been actively used as gene delivery vehicle because of their minimal toxicity, but their relatively low efficiency of gene delivery is the major disadvantage of these vectors. Recently, cysteine residue incorporation to HIV-1 Tat peptide increased liposome-mediated transfection compared with unmodified Tat peptide. Therefore, we designed a novel modified Tat peptide having a homodimeric (Tat-CTHD, Tat-NTHD) and closed structure (cyclic Tat) simply by using the disulfide bond between cysteines to develop a more efficient and safe nonviral gene delivery system. The mixing of Tat-CTHD and Tat-NTHD with DNA before mixing with lipofectamine increased the transfection efficiency compared with unmodified Tat peptide and lipofectamine only in MCF-7 breast cancer cells and rat vascular smooth muscle cells. However, cyclic Tat did not show any improvement in the transfection efficiency. In the gel retardation assay, Tat-CTHD and Tat-NTHD showed more strong binding with DNA than unmodified Tat and cyclic Tat peptide. This enhancement was only shown when Tat-CTHD and Tat-NTHD were mixed with DNA before mixing with lipofectamine. The effects of Tat-CTHD and Tat-NTHD were also valid in the experiment using DOTAP and DMRIE instead of lipofectamine. We could not find any significant cytotoxicity in the working concentration and more usage of these peptides. In conclusion, we have designed a novel transfection-enhancing peptide by easy homodimerization of Tat peptide, and the simple mix of these novel peptides with DNA increased the gene transfer of cationic lipids more efficiently with no additional cytotoxicity.
URI
http://hdl.handle.net/YU.REPOSITORY/24770http://dx.doi.org/10.4014/jmb.1105.05041
ISSN
1017-7825
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의과대학 > 생리학교실 > Articles
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