Department of Medical Biochemistry and Molecular Biology, University of Saarland Medical Center, D-66421 Homburg, Germany.
J Neurosci Methods. 2012;206(2):138-42. doi: 10.1016/j.jneumeth.2012.02.016. Epub 2012 Feb 23.
1321N1 astrocytoma cells are frequently used to analyze stimulus-induced intracellular signaling. These experiments require genetic manipulation of the cells and several chemical and physical methods have been employed in the past. Recently, microporation has been suggested as the best method to transfect 1321N1 astrocytoma cells. Here, we demonstrate that lentiviral gene transfer into 1321N1 cells is highly efficient, cheap and non-toxic. In addition, lentiviral gene transfer efficiently facilitates stable expression of small hairpin RNAs. Finally, lentiviral gene transfer can be used to implant promoter/luciferase reporter genes into the chromatin of the cells, allowing promoter studies using templates that are embedded into the nucleosomal structure of the chromatin.
1321N1 星形细胞瘤细胞常用于分析刺激诱导的细胞内信号转导。这些实验需要对细胞进行基因操作，过去曾采用过几种化学和物理方法。最近，微孔穿孔被认为是转染 1321N1 星形细胞瘤细胞的最佳方法。在这里，我们证明了慢病毒基因转移到 1321N1 细胞中非常高效、廉价且无毒。此外，慢病毒基因转移有效地促进了短发夹 RNA 的稳定表达。最后，慢病毒基因转移可用于将启动子/荧光素酶报告基因植入细胞的染色质中，从而使用嵌入染色质核小体结构中的模板进行启动子研究。
