Cell-based high-throughput screening of cationic polymers for efficient DNA and siRNA delivery
Cell-based high-throughput screening of cationic polymers for efficient DNA and siRNA delivery
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基于细胞的阳离子聚合物高通量筛选，用于高效 DNA 和 siRNA 递送
DOI：
10.1016/j.actbio.2020.08.029
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发表时间：
2020
影响因子：
9.7
通讯作者：
Deng Wenbin
中科院分区：
文献类型：
--
作者：
Wu Yihang;Wang Ling;Xiong Yue;Zhou Quanming;Li Linxian;Chen Guanyu;Ping Yulei;Davidson Gary;Levkin Pavel A.;Gao Liqian;Deng Wenbin
开发能够安全有效地将质粒DNA和siRNA导入细胞的非病毒基因载体对基因治疗具有重要意义。尽管已经付出了大量的努力，但开发合适的、转染效率高、细胞毒性低的基因载体仍是当务之急。为此，我们成功地设计，合成和筛选了120个聚合物（二卤化物和胺之间的亲核取代反应）的库。利用基于细胞的转染筛选试验，测试了120种聚合物以评估它们将DNA和siRNA转运到细胞中的转染效率。我们的结果表明，疏水性修饰可以大大提高阳离子聚合物的转染效率，具有长连接基的聚合物通常表现出更好的转染性能，尤其是具有1，12-二溴十二烷（L3连接基）连接基的聚合物。此外，当聚合物粒径在200 nm左右，zeta电位在+40 mV ~+50 mV范围内时，聚烷基胺具有更好的转染效率。有趣的是，由N15 HL 3制成的聚合物颗粒不仅在HEK 293 T细胞中表现出更好的DNA转染效率，而且在U87 Luc-GFP细胞中表现出更高的siRNA转染效率，以及比Lipofectamine 2000（商业转染试剂之一）低的细胞毒性。因此，我们的研究不仅为基因治疗的进一步评价提供了有希望的基因载体候选物，而且为更好地理解化学结构与基因转染效率之间的关系提供了有价值的见解，以合理设计更好的非病毒基因载体用于基因治疗。
Development of non-viral gene vectors which can efficiently and safely transfect plasmid DNA and siRNA into cells is of great importance for gene therapy. Despite lots of efforts spent, it is still imperative to develop suitable gene vectors with better transfection efficiency and low cytotoxicity. To this end, we successfully designed, synthesized and screened a library of 120 polymers (vianucleophilic substitution reaction between dihalides and amines). With cell-based transfection screening assays, 120 polymers were tested to evaluate their transfection efficiency of transporting DNA and siRNA into cells. Our results indicated that hydrophobic modification could greatly enhance cationic polymers’ transfection efficiency, and polymers with long linkers usually showed better transfection performance, especially for polymers with the linker of 1, 12-dibromododecane (L3 linker). Besides, polyalkylamines exhibited better transfection efficiency with the polymer particle size around 200 nm and the zeta potential in the range of + 40 mV to +50 mV. Interestingly, polymer particles made from N15HL3 not only exhibited better DNA transfection efficiency in HEK 293T cells but also showed higher siRNA transfection efficiency in U87 Luc-GFP cells together with low cell toxicity than Lipofectamine 2000 (one of commercial transfection reagents). Therefore, it is hoped that our study here not only provides promising gene vector candidates for further evaluation in gene therapy, but also provides valuable insights for better understanding of the relationship between the chemical structures and gene transfection efficiency to rationally design better non-viral gene vectors for gene therapy in the future.