Stabilization of poly-L-lysine/DNA polyplexes for in vivo gene delivery to the liver
Stabilization of poly-L-lysine/DNA polyplexes for in vivo gene delivery to the liver
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DOI：
10.1016/s0167-4781(98)00274-7
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发表时间：
1999-02-16
期刊：
影响因子：
--
通讯作者：
Carlo, DJ
中科院分区：
文献类型：
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作者：
Kwoh, DY;Coffin, CC;Carlo, DJ
我们正在开发一种基于聚L-赖氨酸和质粒DNA的自组装非病毒体内基因传递载体，我们已经表征了不同链长度的聚L-赖氨酸的DNA缩合和DNA结合强度。大于 20 个残基的多聚 L-赖氨酸链可以在生理盐水中有效结合 DNA，而较短的链则不能。脱唾液酸类粘蛋白与 PLL 的连接增加了盐水中有效 DNA 结合和有效 DNA 浓缩所需的 PLL 链长度。通过电子显微镜观察，聚-L-赖氨酸/DNA复合物呈直径25-50 nm的环形或40-80 nm长的棒状；脱唾液酸类粘蛋白与聚赖氨酸成分的缀合将所得聚合复合物的尺寸增加至50-90 nmn。在水中，聚-L-赖氨酸和脱唾液酸类粘蛋白-PLL 复合物的有效直径分别为 46 和 87.6 nm。仅含有聚-L-赖氨酸和DNA的聚合复合物在生理盐水中以所有电荷比聚集，并在水中以中性电荷比聚集。脱唾液酸类粘蛋白的附着减少但没有消除PLL复合物的聚集，并且没有导致复合物有效递送至肝细胞。聚乙二醇与聚-L-赖氨酸的缀合通过屏蔽表面以中性电荷比产生空间稳定的复合物。为了有效地体内基因传递，聚合复合物需要空间稳定以防止聚集和与血清成分相互作用。 (C) 1999 年由 Elsevier Science B.V. 出版。保留所有权利。
We are developing a self-assembling non-viral in vivo gene delivery vehicle based on poly-L-lysine and plasmid DNA, We have characterized poly-L-lysines of different chain lengths for DNA condensation and strength of DNA binding. Poly-L-lysine chains > 20 residues bound DNA efficiently in physiological saline, while shorter chains did not. Attachment of asialoorosomucoid to PLL increased the PLL chain length required for efficient DNA binding in saline and for efficient DNA condensation. By electron microscopy, poly-L-lysine/DNA polyplexes appeared as toroids 25-50 nm in diameter or rods 40-80 nm long; conjugation of asialoorosomucoid to the polylysine component increased the size of resulting polyplexes to 50-90 nmn. In water, poly-L-lysine and asialoorosomucoid-PLL polyplexes have effective diameters of 46 and 87.6 nm, respectively. Polyplexes containing only poly-L-lysine and DNA aggregated in physiological saline at all charge ratios and aggregated at neutral charge ratios in water. Attachment of asialoorosomucoid lessened, but did not eliminate, the aggregation of PLL polyplexes, and did not result in efficient delivery of polyplexes to hepatocytes. Conjugation of polyethylene glycol to poly-L-lysine sterically stabilized resulting polyplexes at neutral charge ratios by shielding the surfaces. For efficient in vivo gene delivery, polyplexes will need to be sterically stabilized to prevent aggregation and interaction with serum components. (C) 1999 Published by Elsevier Science B.V. All rights reserved.