[Peptide TAT modified polyethylenimine-beta-cyclodextrin for gene delivery].
To develop a novel gene delivery vector TAT-PEI-beta-CyD.



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.



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.



The novel vector TAT-PEI-beta-CyD has been developed successfully with low cytotoxicity and high transfection efficiency.
Lai
Li-Hua
LH
Institute of Immunology, College of Medicine, Zhejiang University, Hangzhou 310058, China.
Jiang
Qi-Ying
QY
Chen
Dan
D
Hu
Yi-Ping
YP
Yu
Hai
H
Wang
Qing-Qing
QQ
Tang
Gu-Ping
GP
D002460:Cell Line; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D010446:Peptide Fragments / Q000737:chemistry*; D011094:Polyethyleneimine / Q000737:chemistry*; D047392:beta-Cyclodextrins / Q000737:chemistry*; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000737:chemistry*
D010446:Peptide Fragments; D047392:beta-Cyclodextrins; D054322:tat Gene Products, Human Immunodeficiency Virus; D011094:Polyethyleneimine; C031215:betadex