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Journal of Zhejiang University. Medical sciences ; (6): 15-23, 2009.
Article in Chinese | WPRIM | ID: wpr-310396

ABSTRACT

<p><b>OBJECTIVE</b>To develop a novel gene delivery vector TAT-PEI-beta-CyD.</p><p><b>METHODS</b>beta-cyclodextrin (beta-CyD) was linked by low molecular weight (PEI 600) via 1, 1-carbonyldiimidazole (CDI), and TAT peptide (RRRQRRKKRC) was coupled to PEI 600 by [N-succinimidy-3-(2-pyridyldithio) propionate, SPDP]. The copolymer was characterized by (1)H-NMR and FT-IR. Physiochemical characteristics of TAT-PEI-beta-CyD/DNA complexes were tested by agarose gel electrophoresis and particle size measurements. Cell viability and transfection efficiency were evaluated in A293 and B16 cells using PEI 25 kDa as a control.</p><p><b>RESULT</b>TAT peptide was successfully coupled to PEI-beta-CyD. The result of gel electrophoresis showed that the TAT-PEI-beta-CyD was able to condense DNA efficiently at N/P ratio of 4. The particle size of TAT-PEI-beta-CyD/DNA complexes was around 100 nm. The cytotoxicity of TAT-PEI-beta-CyD was lower than that of PEI 25 kDa. The transfection efficiency of TAT-PEI-beta-CyD was higher than that of PEI 25 kDa in A293 and B16 cells at N/P ratio of 30.</p><p><b>CONCLUSION</b>The novel vector TAT-PEI-beta-CyD has been developed successfully with low cytotoxicity and high transfection efficiency.</p>


Subject(s)
Humans , Cell Line , Gene Transfer Techniques , Genetic Therapy , Methods , Peptide Fragments , Chemistry , Polyethyleneimine , Chemistry , beta-Cyclodextrins , Chemistry , tat Gene Products, Human Immunodeficiency Virus , Chemistry
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