Take the 2-minute tour ×
Biology Stack Exchange is a question and answer site for biology researchers, academics, and students. It's 100% free, no registration required.

Does anyone have experience with the crispr/cas9 platform performed on MEF? Or does anyone recall any relevent articles?
Thanks

share|improve this question
2  
Is this the paper you are looking for? genesdev.cshlp.org/content/27/23/2602.long –  Devashish Das Jul 21 at 12:51

1 Answer 1

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.

Furthermore:

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.

share|improve this answer

Your Answer

 
discard

By posting your answer, you agree to the privacy policy and terms of service.

Not the answer you're looking for? Browse other questions tagged or ask your own question.