Combinatorial Approach to Determine Functional Group Effects on Lipidoid-Mediated siRNA Delivery
Combinatorial Approach to Determine Functional Group Effects on Lipidoid-Mediated siRNA Delivery
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DOI：
10.1021/bc100041r
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发表时间：
2010-08-01
影响因子：
4.7
通讯作者：
Anderson, Daniel G.
中科院分区：
文献类型：
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作者：
Mahon, Kerry P.;Love, Kevin T.;Anderson, Daniel G.
RNA干扰（RNAi）的应用，无论是在临床还是在实验室，都需要安全有效的给药方法。在这里，我们开发了一种组合方法来合成一个基于两种具有已知siRNA传递能力的脂质样底物的传递载体库。该文库的成员具有脂质样尾巴和含有羟基，氨基甲酸酯，醚或胺官能团以及烷基链长度和分支变化的特征附属物的混合物。利用荧光素酶报告系统，我们研究了脂质化学修饰与体外递送性能的关系。结果表明，功能基团的影响取决于总胺含量和递送载体的尾部数量。此外，使用因子VII敲除试验评估体内性能。两个文库成员，每个都含有醚基团，被发现在与母体传递载体相当的水平上击倒目标蛋白。这些结果表明，传递载体的微小化学变化会影响体外和体内的敲除效率和细胞活力。本文描述的工作确定了siRNA递送的新材料，并为优化脂质样siRNA递送材料的化学组成参数提供了新的见解。
The application of RNA interference (RNAi), either in the clinic or in the laboratory, requires safe and effective delivery methods. Here, we develop a combinatorial approach to synthesize a library of delivery vectors based on two lipid-like substrates with known siRNA delivery capabilities. Members of this library have a mixture of lipid-like tails and feature appendages containing hydroxyl, carbamate, ether, or amine functional groups as well as variations in alkyl chain length and branching. Using a luciferase reporter system in He La cells, we studied the relationship between lipid chemical modification and delivery performance in vitro. The impact of the functional group was shown to vary depending on the overall amine content and tail number of the delivery vector. Additionally, in vivo performance was evaluated using a Factor VII knockdown assay. Two library members, each containing ether groups, were found to knock down the target protein at levels comparable to those of the parent delivery vector. These results demonstrate that small chemical changes to the delivery vector impact knockdown efficiency and cell viability both in vitro and in vivo. The work described here identifies new materials for siRNA delivery and provides new insight into the parameters for optimized chemical makeup of lipid-like siRNA delivery materials.