Attaching Biosynthesized Bacterial Magnetic Particles to Polyethylenimine Enhances Gene Delivery Into Mammalian Cells
Attaching Biosynthesized Bacterial Magnetic Particles to Polyethylenimine Enhances Gene Delivery Into Mammalian Cells
复制标题
将生物合成的细菌磁性颗粒附着到聚乙烯亚胺上可增强基因向哺乳动物细胞的传递
DOI：
10.1166/jbn.2016.2213
复制
发表时间：
2016
影响因子：
2.9
通讯作者：
Fang Meiying
中科院分区：
文献类型：
--
作者：
Yang Wanjie;Bai Ying;Wang Xu;Dong Xinxing;Li Ying;Fang Meiying
使用细菌磁性颗粒（BMPs）-聚乙烯亚胺（PEI）的基因转染已经变得越来越普遍;然而，相对较少的努力已经被用于优化用于制备这些复合物的方案以提高它们的转染效率。在这里，我们报告了一种用于构建BMP-PEI/DNA复合物的方法，该方法导致转染效率提高，细胞毒性降低，复合物形成和转染的操作时间缩短。使用超声处理混合的BMPs-PEI/DNA复合物产生的珠粒比通过机械涡旋制备的复合物更有效地分离HeLa细胞10.2%。磷酸盐缓冲液（PBS）是BMPs-PEI/DNA和PEI/DNA复合物的上级溶剂，转染率分别为54.75%和46.01%。与用标准PEI/DNA复合物孵育4小时相比，用低剂量BMPs-PEI/DNA复合物孵育10分钟后实现了相当的转染水平。储存在PBS中的BMP具有比储存在生理盐溶液中的BMP高5%的平均转染效率。细胞形态学和细胞毒性分析表明，生物合成的BMP减轻PEI对细胞的毒性。我们的研究结果为体外构建BMPs-PEI/DNA复合物和基因转移提供了一个优化的方案。
Gene transfection using bacterial magnetic particles (BMPs)-polyethylenimine (PEI) has become increasingly prevalent; however, relatively little effort has been made to optimize the protocol for preparing these complexes with the aim of improving their transfection efficiency. Here, we report a procedure for constructing BMPs-PEI/DNA complexes that results in improved transfection efficiency, reduced cytotoxicity and shorter procedure times for both complex formation and transfection over current methods. BMPs-PEI/DNA complexes mixed using ultrasonication yielded beads that were 10.2% more efficient at transfecting HeLa cells than complexes made by mechanical vortexing. Phosphate-buffered saline (PBS) proved to be a superior solvent for BMPs-PEI/DNA and PEI/DNA complexes, and the transfection efficiencies in HeLa cells were 54.75% and 46.01%, respectively. Comparable levels of transfection were achieved after 10 min of incubation with low-dose BMPs-PEI/DNA complexes versus 4 h with standard PEI/DNA complexes. BMPs stored in PBS have an average transfection efficiency that is 5% greater than those stored in physiological salt solutions. Cell morphology and cytotoxicity analyses demonstrated that the biosynthesized BMPs lessened the cytotoxicity of PEI to cells. Our results provide an optimized protocol for BMPs-PEI/DNA complex construction and gene transfer in vitro.