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: Catherine Vaughan, Swati Palit Deb, Sumitra Deb
• Journal: Methods in molecular biology (Clifton, N.J.) (Methods Mol Biol) Vol. 962 Pg. 193-9 ( 2013) ISSN: 1940-6029 [Electronic] United States
• PMID: 23150448 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• DNA, Viral
• Mutant Proteins
• RNA, Messenger
• RNA, Small Interfering
• Tumor Suppressor Protein p53

Topics

• DNA, Viral (genetics)
• Gene Knockdown Techniques (methods)
• Genetic Vectors
• HEK293 Cells
• Humans
• Lentivirus
• Mutant Proteins (genetics)
• RNA, Messenger (genetics)
• RNA, Small Interfering (genetics)
• Transfection
• Tumor Suppressor Protein p53 (genetics)