shRNA screens for mediators of anti-proliferative effects of p53 in SJSA cell line

To identify genes required for anti-proliferative function of p53 we performed genome-wide and p53TARGET screens in SJSA cells treated with Nutlin. Overall design: SJSA cells were transduced with the pooled libraries in triplicate and propagated for at least 7 days. Next, cells were exposed to either DMSO or 10 microM Nutlin for 48 hours. After recovery, cells were treated one more time for 48 hours. Following final recovery period, gDNA was harvested and shRNA expression casettes were amplified by PCR. Purified reaction product was used for preparation of sequencing library.