Rapid Discovery of Potent siRNA-Containing Lipid Nanoparticles Enabled by Controlled Microfluidic Formulation
Rapid Discovery of Potent siRNA-Containing Lipid Nanoparticles Enabled by Controlled Microfluidic Formulation
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DOI：
10.1021/ja301621z
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发表时间：
2012-04-25
影响因子：
15
通讯作者：
Anderson, Daniel G.
中科院分区：
文献类型：
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作者：
Chen, Delai;Love, Kevin T.;Anderson, Daniel G.
用于核酸体内递送的有效新材料的发现取决于将活性分子成功地配制成适合于生理环境的剂型。由于目前的配制方法效率低下，通常首先评估材料作为与核酸的简单离子复合物（脂质复合物）的体外递送功效。然而，这种测定的预测价值从未被系统地研究过。在这里，第一次，通过开发一种微流控方法，允许快速制备高质量的含siRNA的脂质纳米颗粒（LNP）的大量材料，我们已经表明，采用lipoplexes的基因沉默测定导致高假阴性率（类似于90%），这在很大程度上可以通过配制来避免。发现了7种新材料，其在小鼠中以1.0 mg/kg的剂量具有>90%的体内基因沉默效力。该方法将有助于发现用于LNP介导的核酸递送的下一代试剂。
The discovery of potent new materials for in vivo delivery of nucleic acids depends upon successful formulation of the active molecules into a dosage form suitable for the physiological environment. Because of the inefficiencies of current formulation methods, materials are usually first evaluated for in vitro delivery efficacy as simple ionic complexes with the nucleic acids (lipoplexes). The predictive value of such assays, however, has never been systematically studied. Here, for the first time, by developing a microfluidic method that allowed the rapid preparation of high-quality siRNA-containing lipid nanoparticles (LNPs) for a large number of materials, we have shown that gene silencing assays employing lipoplexes result in a high rate of false negatives (similar to 90%) that can largely be avoided through formulation. Seven novel materials with in vivo gene silencing potencies of >90% at a dose of 1.0 mg/kg in mice were discovered. This method will facilitate the discovery of next-generation reagents for LNP-mediated nucleic acid delivery.