How to CRISPR-KO and CRISPR-HDR of p53 in fibroblasts?

16 January 2023 0 8K Report

I performed CRISPR-KO of p53 in fibroblasts. I've got cells that lost the activity of WTp53 (several target genes tested on qPCR) but on western blot, I see still quite a high level of a protein that is slightly lower than WTp53, and is not inducible by nutlin. Any idea how to get rid of these remainings?

The other thing is CRISPR-HDR to introduce gain of function mutations e.g. R175h and R273H. Do you have any advice how to increase the efficiency?

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