Methods in Molecular Biology Volume 962, 2013, pp 193-199
Date: 21 Sep 2012

Generation of p53 Knock-down Cell Lines

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In order to study the functions of a cell’s endogenous mutant p53, the p53 protein levels must be knocked-down. Transient transfection of small interfering RNAs is one way to accomplish this. Another is the stable expression of short hairpin RNAs. This chapter presents a method by which a short hairpin RNA (shRNA) targeting p53 is inserted into the genome of a cell via lentivirus infection. These p53 knock-down cell lines are stable and may be grown long term for use in a wide range of applications.