p53 knockdown - (Aug/19/2008 )

Hi all,

I have never worked with projects that involve silencing of any gene. I am now interested in silencing the p53 gene in a human cell line. Does anyone have the protocol for the same or if someone could guide me in the right direction.

thanks

pooja

The protocol is the same as any transfection if you are using oligos (i.e not virus) to perform the knockdown. You will need to titrate the most effective dosage which also has the least non-specific effect on the cells, this will usually be in the low nanomol per litre range.

For getting siRNA etc there are a few databases around, just have a google for them. Also have a look at papers and try the published sequences.

You can get a pSico plasmid (lentivirus ready) with p53 shRNA from Addgene.com at very low cost. This is the 1 developed by Tyler Jacks. Or you can just find published p53 siRNAs as suggested by Bob.

thanks a bunch,

I will look around in the databases. I was not aware that those existed.

Pooja

thanks a bunch,

I will try the pSico plasmid since I am also using the mouse genotyping protocol developed in Tyler Jacks' lab.

Pooja