Stability of peptide condensed plasmid DNA formulations
Stability of peptide condensed plasmid DNA formulations
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DOI：
10.1021/js9800477
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发表时间：
1998-06-01
影响因子：
3.8
通讯作者：
Rice, KG
中科院分区：
文献类型：
--
作者：
Adami, RC;Collard, WT;Rice, KG
低分子量均质肽用于形成肽/DNA缩合物。具有18个赖氨酸的肽被发现保护质粒DNA免受血清内切酶和超声诱导的降解，而具有8个赖氨酸的较短的肽在0.1 M氯化钠中解离，未能保护DNA免受酶促降解。在100 W超声处理长达60 s后，肽浓缩的DNA显示超螺旋与环状DNA的比率没有变化，并且能够以与未处理的浓缩质粒DNA相同的效率转染HepG 2细胞。或者，未浓缩的质粒DNA通过超声处理和血清核酸内切酶迅速片段化，并在与肽浓缩后导致可忽略的基因表达。阳离子脂质/DNA复合物在稳定血清中的DNA方面仅部分有效，而肽/DNA缩合所提供的完全稳定则相反。这些结果表明，通过与肽缩合提供的稳定性在配制期间保护DNA并在血清中保持其结构。这些功能对于从非病毒基因递送系统实现最佳基因表达是重要的。
Low molecular weight homogeneous peptides were used to form peptide/DNA condensates. A peptide possessing 18 lysines was found to protect plasmid DNA from serum endonuclease and sonicative-induced degradation whereas a shorter peptide possessing 8 lysines dissociated in 0.1 M sodium chloride and failed to protect DNA from enzymatic degradation. Peptide-condensed DNA showed no change in the ratio of supercoiled to circular DNA following 100 W sonication for up to 60 s and was able to transfect HepG2 cells with equivalent efficiency as untreated condensed plasmid DNA. Alternatively, uncondensed plasmid DNA was rapidly fragmented by sonication and serum endonucleases and resulted in negligible gene expression following condensation with peptide. Cationic lipid/DNA complexes were only partially effective at stabilizing DNA in serum compared to the complete stabilization afforded by peptide/DNA condensation. These results indicate that the stabilization afforded by condensation with a peptide protects DNA during formulation and preserves its structure in serum. These functions are important to achieve optimal gene expression from a nonviral gene delivery system.