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Generation of p53 Knock-down Cell Lines

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p53 Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 962))

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.

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References

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Acknowledgments

This work was supported by grants from NIH to Sumitra Deb (CA70712 and CA121144) and Swati Palit Deb (CA74172), and Pilot Project Awards from Massey Cancer to Sumitra Deb and Swati Palit Deb. We thank Shilpa Singh for help in editing this chapter.

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Authors and Affiliations

  1. Department of Integrative Life Sciences, Virginia Commonwealth University, Richmond, VA, USA

    Catherine Vaughan

  2. Department of Biochemistry, Massey Cancer Center, Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond, VA, USA

    Swati Palit Deb & Sumitra Deb

Authors
  1. Catherine Vaughan
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  2. Swati Palit Deb
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  3. Sumitra Deb
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Corresponding author

Correspondence to Sumitra Deb .

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Editors and Affiliations

  1. Massey Cancer Center, Department of Biochemistry, Virginia Commonwealth University, College Street 401, Richmond, 23298, Virginia, USA

    Sumitra Deb

  2. Massey Cancer Center, Department of Biochemistry, Virginia Commonwealth University, College Street 401, Richmond, 23298, Virginia, USA

    Swati Palit Deb

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© 2013 Springer Science+Business Media New York

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Vaughan, C., Deb, S.P., Deb, S. (2013). Generation of p53 Knock-down Cell Lines. In: Deb, S., Deb, S. (eds) p53 Protocols. Methods in Molecular Biology, vol 962. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-236-0_16

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  • DOI: https://doi.org/10.1007/978-1-62703-236-0_16

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-235-3

  • Online ISBN: 978-1-62703-236-0

  • eBook Packages: Springer Protocols

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