Peptide TAT modified polyethylenimine-beta-cyclodextrin for gene delivery.
- Author:
Li-Hua LAI
1
;
Qi-Ying JIANG
;
Dan CHEN
;
Yi-Ping HU
;
Hai YU
;
Qing-Qing WANG
;
Gu-Ping TANG
Author Information
- Publication Type:Journal Article
- MeSH: Cell Line; Gene Transfer Techniques; Genetic Therapy; methods; Humans; Peptide Fragments; chemistry; Polyethyleneimine; chemistry; beta-Cyclodextrins; chemistry; tat Gene Products, Human Immunodeficiency Virus; chemistry
- From: Journal of Zhejiang University. Medical sciences 2009;38(1):15-23
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo develop a novel gene delivery vector TAT-PEI-beta-CyD.
METHODSbeta-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.
RESULTTAT 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.
CONCLUSIONThe novel vector TAT-PEI-beta-CyD has been developed successfully with low cytotoxicity and high transfection efficiency.
