Liu Dan, Songyang Zhou
Cell-based Assay Screening Service Core, Baylor College of Medicine, Houston, Texas, USA.
Methods Enzymol. 2008;446:409-19. doi: 10.1016/S0076-6879(08)01624-8.
Genetic screens have been proven powerful for the identification of components of various signaling pathways. For mammalian cells, methods for genetic screens are limited. We have developed the ERM (enhanced retroviral mutagen) mutagenesis approach that has been shown to be efficient and amenable to genomewide genetic screens in mammalian cells without the need of cDNA library construction. The ERM method offers several advantages, including conditional gene expression and the flexibility to tag endogenous genes with different epitope-tag and marker sequences. This chapter will discuss general design, procedures, and applications of the ERM strategy.
基因筛选已被证明在识别各种信号通路的组成部分方面非常有效。对于哺乳动物细胞,基因筛选方法有限。我们开发了ERM(增强逆转录病毒诱变)诱变方法,该方法已被证明是有效的,并且适用于哺乳动物细胞的全基因组遗传筛选,而无需构建cDNA文库。ERM方法具有几个优点,包括条件性基因表达以及用不同表位标签和标记序列标记内源基因的灵活性。本章将讨论ERM策略的一般设计、程序和应用。