Physical properties and in vitro transfection efficiency of gene delivery vectors based on complexes of DNA with synthetic polycations
Physical properties and in vitro transfection efficiency of gene delivery vectors based on complexes of DNA with synthetic polycations
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DOI：
10.1016/s0168-3659(02)00045-7
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发表时间：
2002-05-17
影响因子：
10.8
通讯作者：
Ulbrich, K
中科院分区：
文献类型：
--
作者：
Reschel, T;Konák, C;Ulbrich, K
研究了用于基因递送的聚阳离子/DNA复合物的生物物理性质，包括其制备条件、所涉及的聚阳离子的化学结构和分子量。所使用的聚阳离子包括各种含有伯氨基和叔氨基或季铵基团的阳离子聚合物和共聚物。研究发现，这些聚电解质复合物 (PEC) 的分子量和尺寸随着缓冲液温度和 pH 值的增加而增加。通过降低用于形成PEC的聚阳离子的分子量，复合物在较低pH值的水溶液中变得不稳定，难以凝固。DNA与低分子量聚阳离子在水中的自组装为PEC提供了最低的分子量、最小的尺寸和最低的密度，但它们在氯化钠溶液中的稳定性非常差。尽管多步转染过程很复杂。发现体外转染效率与复合物在 NaCl 溶液中的稳定性以及在 0.15 M NaCl 溶液中的凝固之间存在直接相关性。含有伯氨基的聚阳离子的DNA复合物在盐溶液中表现出最好的稳定性，并且也表现出最好的转染活性。由具有季铵基团的聚阳离子形成的PEC对所添加的盐的破坏的抵抗力最差，并且在转染测定中提供最低的活性。发现用含有伯胺和叔胺的统计共聚物形成的 DNA 复合物具有最高的转染活性。 (C) 2002 Elsevier Science B.V. 保留所有权利。
Biophysical properties of polycation/DNA complexes designed for gene delivery were studied with respect to the conditions of their preparation, chemical structure and molecular weight of the polycations involved. The polycations used included a variety of cationic polymers and copolymers containing primary and tertiary amino or quaternary ammonium groups. It was found that the molecular weight and the size of these polyelectrolyte complexes (PECs) increase with increasing temperature and pH of the buffer. By decreasing the molecular weight of polycations used for PEC formation, the complexes become unstable towards coagulation in aqueous solution at lower pH, The self-assembly of DNA with low-molecular-weight polycations in water provides PECs with the lowest molecular weight, smallest size and the lowest density but their stability in NaCl solutions is very poor. Despite the complexity of the multistep transfection process. a direct correlation between the transfection efficiency in vitro and the stability of the complexes in NaCl solutions and coagulation in 0.15 M NaCl solution was found. DNA complexes with polycations containing primary amino groups showed the best stability in saline solutions and also the best transfection activity. PECs formed by polycations with quaternary ammonium groups were the least resistant to destruction by the added salt and provided the lowest activity in transfection assays. The highest transfection activity was found for DNA complexes formed with a statistical copolymer containing primary and tertiary amines. (C) 2002 Elsevier Science B.V. All rights reserved.