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Related Concept Videos

In-vitro Mutagenesis01:16

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
Abnormal Proliferation02:23

Abnormal Proliferation

Under normal conditions, most adult cells remain in a non-proliferative state unless stimulated by internal or external factors to replace lost cells. Abnormal cell proliferation is a condition in which the cell's growth exceeds and is uncoordinated with normal cells. In such situations, cell division persists in the same excessive manner even after cessation of the stimuli, leading to persistent tumors. The tumor arises from the damaged cells that replicate to pass the damage to the daughter...

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Related Experiment Video

Updated: May 16, 2026

Generation of Stable Human Cell Lines with Tetracycline-inducible (Tet-on) shRNA or cDNA Expression
09:51

Generation of Stable Human Cell Lines with Tetracycline-inducible (Tet-on) shRNA or cDNA Expression

Published on: March 5, 2013

Generation of p53 knock-down cell lines.

Catherine Vaughan1, Swati Palit Deb, Sumitra Deb

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

Methods in Molecular Biology (Clifton, N.J.)
|November 15, 2012
PubMed
Summary

Researchers developed a method to stably reduce p53 protein levels using lentivirus-delivered short hairpin RNA (shRNA). This creates long-lasting cell lines for studying mutant p53 functions in various applications.

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Last Updated: May 16, 2026

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09:51

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13:54

Preparation of Cell-lines for Conditional Knockdown of Gene Expression and Measurement of the Knockdown Effects on E4orf4-Induced Cell Death

Published on: October 21, 2012

Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Understanding the function of endogenous mutant p53 requires methods to reduce its protein levels.
  • Both transient small interfering RNAs (siRNAs) and stable short hairpin RNAs (shRNAs) can achieve p53 knockdown.
  • Existing methods may have limitations for long-term studies.

Purpose of the Study:

  • To present a reliable method for creating stable p53 knockdown cell lines.
  • To facilitate the study of endogenous mutant p53 protein functions.
  • To establish cell lines suitable for long-term research applications.

Main Methods:

  • Utilizing lentivirus infection to deliver a short hairpin RNA (shRNA) targeting p53.
  • Integrating the shRNA construct into the host cell genome for stable expression.
  • Establishing and maintaining long-term cell cultures derived from the infected cells.

Main Results:

  • Successful generation of stable p53 knockdown cell lines.
  • Demonstration of long-term p53 protein level reduction.
  • Cell lines are viable for extended culture periods.

Conclusions:

  • Lentiviral delivery of shRNA provides a robust method for stable p53 knockdown.
  • These engineered cell lines are valuable tools for investigating p53 biology.
  • The method supports diverse research applications requiring sustained p53 reduction.