Studies on the interactions between DNA and PAMAM with fluorescent probe [Ru(phen)2 dppz]2+.
The interactions between plasmid DNA and cationic polymers are of interest for their potential biological applications. In this paper, the interactions of DNA with polyamidoamine (PAMAM) dendrimer were studied by fluorescence spectroscopy and stopped-flow technique. A rapid and reproducible fluorescent assay method had been developed for assessing PAMAM and DNA interactions using [Ru(phen)2 dppz]2+ as a probe. We further studied the kinetics of PAMAM binding to DNA and the reverse process of DNA dissociation from the complexes. The results indicated that DNA condensation was the rate-determining step during the complexation process, while DNA unfolding and expansion was the rate-determining step during the DNA dissociation process. At N/P ratios before reaching the thermodynamically most stable state, the complexes of DNA and PAMAM were incompact and could dissociate to some extent. And at N/P ratios above 2.0, DNA was fully condensed by PAMAM and dissociation was increasingly difficult. These results provided some useful instructions for self-assembling and disassembling of DNA as well as efficient gene delivery applications.
Zhou
Linzhu
L
College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, PR China.
Gan
Lu
L
Li
Haibing
H
Yang
Xiangliang
X
D006690:Bisbenzimidazole; D004396:Coloring Agents; D004247:DNA / Q000737:chemistry*; D050091:Dendrimers; D004996:Ethidium; D005456:Fluorescent Dyes; D007211:Indoles; D007364:Intercalating Agents; D007700:Kinetics; D049449:Luminescence; D009942:Organometallic Compounds / Q000737:chemistry*; D010777:Photochemistry; D010957:Plasmids / Q000737:chemistry; D011073:Polyamines / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D013816:Thermodynamics
D004396:Coloring Agents; D050091:Dendrimers; D005456:Fluorescent Dyes; D007211:Indoles; D007364:Intercalating Agents; D009942:Organometallic Compounds; C104700:PAMAM Starburst; D011073:Polyamines; C466600:Ru(phen)(2)dipyrido(3,2-a-2',3'-c)phenazine; C007293:DAPI; D004247:DNA; D004996:Ethidium; D006690:Bisbenzimidazole