Abstract：
Isogenic H/N/KRAS-less mouse embryonic fibroblast (MEF) cell lines have been developed to assist in cell-based assays of RAS inhibitors. The quality control assessment of a panel of these isogenic MEFs is described here, with a focus on ensuring the proper insertion of the desired mutant RAS transgene, a determination of gene copy number, and an investigation of potential off-target mutations which could lead to phenotypes which are undesired in downstream experiments. Using this suite of quality control tools, a MEF cell line can be readily validated, and researchers can be assured of the rationale for an observed phenotype.
摘要：
已开发了无等基因H/N/KRAS的小鼠胚胎成纤维细胞(MEF)细胞系以辅助RAS抑制剂的基于细胞的测定。这里描述了一组等基因MEF的质量控制评估，专注于确保正确插入所需的突变RAS转基因，基因拷贝数的测定，以及对可能导致下游实验中不希望出现的表型的潜在脱靶突变的研究。使用这套质量控制工具，MEF细胞系可以很容易地验证，研究人员可以确信观察到的表型的基本原理。
