Any apoptosis experts in the house?

[Hercules Rockefeller]

I have a large pile of papers on my desk that will probably answer my question, but it’s also good to get some personal opinions from experienced people, as I do not have much apoptosis experience or knowledge myself.

My question is: Is there a “best” way to induce apoptosis? I am specifically referring to apoptosis via the intrinsic cell autonomous mechanisms (ie. mitochondrial pathway associated mechanisms) and not any of the extrinsic death receptor mediated mechanisms (ie. Fas ligand, TNF etc).

In other words, overexpression of what intracellular molecule is best at inducing apoptosis?

-- p53 ?

-- Bax ?

-- cytochrome c ?

-- Bcl-2 ?

-- Apaf-1 ?

-- any of the activated caspases?

Seeing as activated caspases are downstream of those other apoptosis–related molecules, is it best to express one of them? If so, which one? Caspase 9? Caspase 3?

Something to note is that I am not talking about cell culture experiments but <I>in vivo</I> experiments in transgenic animals. I am hatching a plan to make transgenic animals that express pro-apoptotic molecules under control of a tissue specific promoter. So, when the promoter becomes active, those specific cells express the prop-apoptotic molecule and die.

Any thoughts?

Thanks.

 

[bmengels]

caspases are activated by digestio, right? so if you know what the activated sequence (protein) of a relevant caspase is, you could make an expression construct for the truncated=activated caspase.
im not even remotely an expert on apoptosis, but i can imagine that different cell-types would be differentially sensitive to the various factors. check whether tha factor you would want to use would be effective in your target tissue. (test in vitro?)
good luck; nice project
BME