Center of Clinical Laboratory, Zhongshan Hospital, Medical College of Xiamen University, Xiamen 361004, China.
J Cell Biochem. 2010 Nov 1;111(4):881-8. doi: 10.1002/jcb.22776.
To synthesize a lipid-cationic polymer (LCP) containing brassidic acid side chain and to investigate its transfection efficiency and characteristics as a siRNA gene vector. The LCP was chemically synthesized and its nucleic acid binding capacity was determined by gel electrophoresis. HeLa-EGFP and TH1080-EGFP cell lines were transfected with siRNA against enhanced green fluorescent protein (EGFP) gene using a LCP to investigate the transfection efficiency. An MTT assay was performed to evaluate the cellular toxicity of the LCP vector. Its degradability and stability under acidic conditions were also investigated. The LCP vector possessed high DNA binding capacity. More than 73% of the cellular fluorescence was inhibited by the LCP-mediated transfection of siRNA against EGFP gene, indicating that vector had high transfection efficiency. Cellular viability was about 95% at the optimum transfection efficiency of LCP, suggesting that the cellular toxicity of LCP was very low. The LCP was also observed to be degradable; moreover, it could be easily stored at normal temperature. A gene vector used for the transfection of siRNA was successfully fabricated from synthesized LCP. Its numerous excellent properties entitle values for further scientific research.
合成一种含有 brassidic 酸侧链的脂阳离子聚合物(LCP),并研究其作为 siRNA 基因载体的转染效率和特性。通过凝胶电泳测定 LCP 的核酸结合能力。用 LCP 转染针对增强型绿色荧光蛋白(EGFP)基因的 siRNA,研究转染效率。通过 MTT 测定法评估 LCP 载体的细胞毒性。还研究了其在酸性条件下的降解性和稳定性。LCP 载体具有高的 DNA 结合能力。LCP 介导的针对 EGFP 基因的 siRNA 转染可抑制超过 73%的细胞荧光,表明载体具有高转染效率。在 LCP 的最佳转染效率下,细胞活力约为 95%,表明 LCP 的细胞毒性非常低。还观察到 LCP 可降解;此外,它可以在常温下轻松储存。成功地从合成的 LCP 制备了用于 siRNA 转染的基因载体。其众多优异的性能为进一步的科学研究提供了价值。