Generation of p53 knock-down cell lines

Methods Mol Biol. 2013:962:193-9. doi: 10.1007/978-1-62703-236-0_16.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 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*

Substances

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