title: Generation of p53 knock-down cell lines.
abstract: 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.
authors.affiliation: Department of Integrative Life Sciences, Virginia Commonwealth University, Richmond, VA, USA.
authors.forename: Catherine
authors.initials: C
authors.lastname: Vaughan
authors.forename: Swati Palit
authors.initials: SP
authors.lastname: Deb
authors.forename: Sumitra
authors.initials: S
authors.lastname: Deb
mesh_terms: D004279:DNA, Viral / Q000235:genetics; D055785:Gene Knockdown Techniques / Q000379:methods*; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus; D050505:Mutant Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000235:genetics*
chemical_list: D004279:DNA, Viral; D050505:Mutant Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D016159:Tumor Suppressor Protein p53