I am designing a CRISPR based assay and need help designing a positive control. Specifically, can someone tell me a protein which causes cell death when it is expressed in cancer cells ? I prefer proteins that do not need post-translational modifications or any kind of activation by other proteins.
Any help is much appreciated. Thank you.
You can go ahead and transfect your cells with PUMA (p53 Upstream Modifier of Apoptosis) and do not treat the cells after transfection with any kind of caspase/pan-caspase inhibitors. You should get huge amount of apoptosis in 24-48 h.
You might try bax protein a pro-apoptotic protein that inhibit the activity of bcl2.
There are so many proteins that causes apoptosis. Out of 25 BCl2 group of proteins some of them are pro-aoptotic (BAX, Bid, BAK, etc) and some of them are anti-apoptotic like (BCl2, BCl-XL, BCl-X) etc. You can use them. Also you can use NOXA, PUMA, p53 etc to check pro-apoptotic activity.
Just overexpress p53 do not make your cells undergo apoptosis, you have to give some kind of stimulus for this. But if you are going to use something to stimulate apoptosis you don`t need p53 overexpression, just to use a chemotherapic as Cisplatin or Doxorrubicin. I would try Bax, as said above.