Impact of Cationic Charge Density and PEGylated Poly(Amino Acid) Tercopolymer Architecture on Their Use as Gene Delivery Vehicles. Part 2: DNA Protection, Stability, Cytotoxicity, and Transfection Efficiency.
Impact of Cationic Charge Density and PEGylated Poly(Amino Acid) Tercopolymer Architecture on Their Use as Gene Delivery Vehicles. Part 2: DNA Protection, Stability, Cytotoxicity, and Transfection Efficiency.
复制标题
阳离子电荷密度和聚乙二醇化聚氨基酸三元共聚物结构对其用作基因递送载体的影响。
DOI：
10.1002/mabi.201800109
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发表时间：
2018
影响因子：
4.6
通讯作者：
Scholz,Carmen
中科院分区：
文献类型：
--
作者：
Ulkoski,David;Scholz,Carmen
研究了PEG‐b‐p(l‐Lys)x‐b‐p(l‐Leu)y、PEG‐b‐p(l‐Leu)x‐b‐p(l‐Lys)y和PEG‐b‐p((l‐Leu)x‐co‐（l‐Lys）y的分子结构对聚乙二醇化聚氨基酸嵌段共聚物转染效率的影响。p（l‐Lys）和p（l‐Leu）的片段长度在10、20和40之间变化；和10和20，分别研究离子/疏水平衡的影响。结果表明，与AB二嵌段共聚物相比，ABC三嵌段共聚物与质粒DNA形成更小、更稳定的多聚体——经长期聚集和溴化乙啶排除研究证实——可以更有效地保护DNA免受核酸酶的侵害，并提供更好的转染效率，如总蛋白和荧光素酶表达所示。更详细的研究表明，与p（l - Leu）形成C -嵌段的三嵌段共聚物在DNA络合中最有效，转染率高2.3倍。此外，通过增加p（l - Lys）链长度来增加阳离子特性，可使转染量提高25%，但同时也会引起一定的细胞毒性。双嵌段共聚物，其中氨基酸构建块作为随机共聚物存在，与DNA结合更松散，导致聚集体不太紧密和不稳定，转染效率较低。
The impact of the molecular architecture on the transfection efficiency of PEGylated poly(amino acid) block copolymers was investigated for PEG‐b‐p(l‐Lys)x‐b‐p(l‐Leu)y, PEG‐b‐p(l‐Leu)x‐b‐p(l‐Lys)y, and PEG‐b‐p((l‐Leu)x‐co‐(l‐Lys)y). The block lengths of p(l‐Lys) and p(l‐Leu) were varied between 10, 20, and 40; and 10 and 20, respectively, to study the influence of the ionic/hydrophobic balance. The results show that ABC triblock copolymers form smaller and more stable polyplexes with plasmid DNA than AB diblock copolymers—as verified by long‐term aggregation and ethidium bromide exclusion studies—protect the DNA more effectively against nucleases, and provide better transfection efficiencies, as indicated by total protein as well as luciferase expression. More detailed studies revealed that triblock copolymers with p(l‐Leu) forming the C‐block were most efficient in DNA complexation with a 2.3 times higher transfection rate. Furthermore, increasing the cationic character by increasing the p(l‐Lys) chain length led to up to 25% higher transfection but at the same time induced some cytotoxicity. Diblock copolymers, where the amino acid–building blocks exist as a random copolymer, bind more loosely with DNA leading to less compact and less stable aggregates with lower transfection efficiencies.