Substrate-mediated DNA delivery: role of the cationic polymer structure and extent of modification
Substrate-mediated DNA delivery: role of the cationic polymer structure and extent of modification
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DOI：
10.1016/j.jconrel.2003.08.003
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发表时间：
2003-11-18
影响因子：
10.8
通讯作者：
Shea, LD
中科院分区：
文献类型：
--
作者：
Segura, T;Volk, MJ;Shea, LD
将DNA复合物固定到支持细胞粘附的基底上可以通过将DNA保持在细胞环境内同时限制复合物聚集来增强基因转移。本报告检查了系绳设计（例如，官能化程度）和阳离子聚合物结构对复合物与底物结合和细胞转染的影响。DNA与阳离子聚合物（聚赖氨酸，PL;聚乙烯亚胺，PEI）复合，所述阳离子聚合物用生物素官能化以结合中性抗生物素蛋白（NA）底物。PL的表面密度为0.4 ~ 2.6 μ g DNA/cm（2），PEI的表面密度为0.7 ~ 1.0 μ g DNA/cm（2）。PL/DNA复合物中生物素基团的分布对细胞转染有双重影响。增加PL与生物素残基的分数降低荧光素酶活性;然而，增加每个PL的生物素残基的数目增加荧光素酶活性。对于PEI，复合物上存在的生物素基团的数量不影响转基因表达。释放研究表明，20-30%的固定化DNA在8天内释放，8-20%在前24小时释放。阳离子聚合物的酶促降解对于转染不是必需的。此外，相对于推注递送，表面介导的递送延长了转基因表达的持续时间。(C)2003 Elsevier B. V.保留所有权利。
DNA complex immobilization to substrates that support cell adhesion can enhance gene transfer by maintaining DNA within the cellular environment while limiting complex aggregation. This report examines the tether design (e.g., extent of functionalization) and cationic polymer structure for their effect on complex binding to the substrate and cellular transfection. DNA is complexed with cationic polymers (polylysine, PL; polyethylenimine, PEI), which are functionalized with biotin for binding to a neutravidin (NA) substrate. Surfaces densities ranging from 0.4 to 2.6 mug DNA/cm(2) were obtained for PL, and from 0.7 to 1.0 mug DNA/cm(2) for PEI. The distribution of biotin groups for PL/DNA complexes had a dual effect on cellular transfection. Increasing the fraction of PL with biotin residues decreased luciferase activity; however, increasing the number of biotin residues per PL increased luciferase activity. For PEI, the number of biotin groups present on the complex did not affect transgene expression. Release studies demonstrated that 20-30% of the immobilized DNA was released over 8 days, with 8-20% released during the first 24 h. Enzymatic degradation of cationic polymers is not necessary for transfection. Additionally, the duration of transgene expression was extended for surface-mediated delivery relative to bolus delivery. (C) 2003 Elsevier B.V. All rights reserved.