Calcium enhances the transfection potency of plasmid DNA-cationic liposome complexes

被引:56
作者
Lam, AMI
Cullis, PR
机构
[1] Univ British Columbia, Fac Med, Dept Biochem & Mol Biol, Vancouver, BC V6T 1Z3, Canada
[2] Inex Pharmaceut Corp, Burnaby, BC V5J 5J8, Canada
来源
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES | 2000年 / 1463卷 / 02期
基金
英国医学研究理事会;
关键词
cationic liposome; plasmid DNA-cationic lipid complex; calcium; transfection;
D O I
10.1016/S0005-2736(99)00219-9
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
It is shown that calcium increases the in vitro transfection potency of plasmid DNA-cationic liposome complexes from 3- to 20-fold. The effect is Ca2+ specific as other cations, such as Mg2+ and Nai, do not give rise to enhanced transfection and the effect can be inhibited by the presence of EGTA. It is shown that Ca2+ increases cellular uptake of the DNA-lipid complexes, indicating that increased transfection potency arises from increased intracellular delivery of both cationic lipid and plasmid DNA in the presence of Ca2+. In particular, it is shown that the levels of intact intracellular plasmid DNA are significantly enhanced when Ca2+ is present. The generality of the Ca2+ effect for enhancing complex-mediated transfection is demonstrated for a number of different cell lines and different cationic lipid formulations. It is concluded that addition of Ca2+ represents a simple and useful protocol for enhancing in vitro transfection properties of plasmid DNA-cationic lipid complexes. (C) 2000 Published by Elsevier Science B.V. All rights reserved.
引用
收藏
页码:279 / 290
页数:12
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