基因转染试剂在NG108-15、SH-SY5Y和CHO-K1细胞系中的转染效率。

E Martín-Montañez, J F López-Téllez, M J Acevedo, J Pavía, Z U Khan
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引用次数: 12

摘要

分析了几种基因传递试剂的转染效率。研究的基因属于哺乳动物g蛋白信号调节因子(regulatory of G-protein signaling, RGS),长度可达2.2 Kb,被转染到NG108-15、SH-SY5Y和CHO-K1细胞系中。转染前,将基因克隆到非病毒载体pcDNA 6.2/EmGFP中,在3′端表达绿色荧光蛋白标签。流式细胞术分析细胞荧光活性,从而分析转染效率。基因传递试剂Lipofectamine 2000和ExGen 500在NG108-15细胞中转染效果更好,而Lipofectamine 2000、ExGen 500和TurboFectin 8.0在CHO-K1细胞中转染效果更好。在这两种细胞系中,转染效率均达到60-80%。在SH-SY5Y细胞中,TurboFectin 8.0转染效果最好;而效率水平仅为5%。基因大小对转染效率无影响。与Lipofectamine 2000不同,使用ExGen 500转染的细胞显示形态变形。结果表明Lipofectamine 2000是最适合转染NG108-15和CHO-K1细胞的转染培养基。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Efficiency of gene transfection reagents in NG108-15, SH-SY5Y and CHO-K1 cell lines.

Several gene delivery reagents were analyzed for their transfection efficiency. Genes studied belonged to the class of mammalian proteins termed regulators of G-protein signaling (RGS), ranged in size up to 2.2 Kb long and were transfected into the NG108-15, SH-SY5Y and CHO-K1 cell lines. Prior to transfection, genes were cloned into a nonviral vector pcDNA 6.2/EmGFP, so as to express a green fluorescent protein tag at the 3' end. Flow cytometry was used to analyze cell fluorescent activity and thereby transfection efficiency. Gene delivery reagents Lipofectamine 2000 and ExGen 500 produced more effective transfection in NG108-15 cells whereas Lipofectamine 2000, ExGen 500 and TurboFectin 8.0 were more effective in CHO-K1 cells. In both these cell lines, transfection efficiency reached 60-80%. In SH-SY5Y cells, TurboFectin 8.0 produced the best transfection result; however efficiency level was only 5%. Gene size had no effect on transfection efficiency. Unlike Lipofectamine 2000, cells transfected using ExGen 500 showed morphological deformation. Our results suggest that Lipofectamine 2000 is the most suitable transfection medium for gene delivery to NG108-15 and CHO-K1 cells.

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