Does anyone have experience with the CRISPR/CAS9 platform performed on MEF? Or does anyone recall any relevant articles?
From this paper:
We chose to test for Cas9-driven targeted mutagenesis on the endogenous Trp53 gene in mouse embryonic fibroblasts (MEFs), a cell line in which the biology of p53 has been thoroughly characterized and where the gene is structurally intact.
Seventy-two hours post-transduction, cells were cultured in the presence or absence of Nutlin-3a, a specific inhibitor of the MDM2–p53 interaction (Tovar et al. 2006). Cells were scored for the fraction of GFP+ cells after 4 d of Nutlin-3a exposure using flow cytometry (Fig. 2B). Nutlin-3a potently activates a p53-dependent anti-proliferative cellular response, which strongly and specifically selects for any cells with disrupted Trp53 function (Efeyan et al. 2007). Much like MLP-p53.1224 infected cells, the pQCiG-p53 GFP+ cells were rapidly enriched for in the presence of Nutlin-3a, suggesting that CRISPR-mediated gene disruption had initially occurred in at least a proportion of the initial Cas9-expressing cells.