Culture of Drosophila S2 cells and their use for RNAi-mediated loss-of-function studies and immunofluorescence microscopy
Culture of Drosophila S2 cells and their use for RNAi-mediated loss-of-function studies and immunofluorescence microscopy
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DOI：
10.1038/nprot.2008.18
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发表时间：
2008-01-01
期刊：
影响因子：
14.8
通讯作者：
Rogers, Gregory C.
中科院分区：
文献类型：
--
作者：
Rogers, Stephen L.;Rogers, Gregory C.
培养的果蝇细胞系已成为细胞生物学和功能基因组学研究中越来越受欢迎的模型系统。最常用的细胞系之一S2细胞特别有用，因为它易于在实验室中生长和维持，对使用RNAi的基因抑制非常敏感，并且非常适合高分辨率光学显微镜测定。在这里，我们提供了S2细胞的常规培养和RNAi处理的方案以及制备这些细胞用于荧光显微镜的方法。使用这些技术，可以在RNAi介导的蛋白质耗尽4-7天后进行功能丧失实验。
Cultured Drosophila cell lines have become an increasingly popular model system for cell biological and functional genomic studies. One of the most commonly used lines, S2 cells, is particularly useful as it is easy to grow and maintain in the lab, is highly susceptible to gene inhibition using RNAi and is well suited to high-resolution light microscopic assays. Here, we provide protocols for the routine culture and RNAi treatment of S2 cells and methods to prepare these cells for fluorescence microscopy. Using these techniques, loss-of-function experiments may be performed after 4-7 d of RNAi-mediated protein depletion.