Related Experiment Video

Updated: Aug 9, 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

Construction by gene targeting in human cells of a "conditional' CDC2 mutant that rereplicates its DNA

J E Itzhaki1, C S Gilbert, A C Porter

  • 1MRC Clinical Sciences Centre, Royal Postgraduate Medical School, Hammersmith Hospital, London, UK.

Nature Genetics
|March 1, 1997
PubMed

Insights

Scientists developed a new gene targeting method to study essential genes. This approach revealed that the CDC2 gene controls DNA replication, preventing premature S-phase entry in human cells.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • Essential genes are crucial for cell survival and function.
  • Understanding cell cycle control is vital for comprehending cell proliferation and disease.
  • The CDC2 gene plays a known role in cell cycle regulation.

Purpose of the Study:

  • To introduce a novel gene targeting strategy for analyzing essential genes in mammalian cells.
  • To investigate the specific function of the cell cycle control gene CDC2 in human cells.
  • To uncover previously unknown control mechanisms in mammalian cell cycle progression.

Main Methods:

  • Development of a novel gene targeting strategy for essential gene analysis.
  • Generation of a specific cell line (HT2-19) where CDC2 expression is inducer-dependent.
  • Observation of cellular responses, including DNA rereplication and apoptosis, in the absence of the inducer.

Main Results:

  • The HT2-19 cell line demonstrated that viability and CDC2 gene expression are dependent on an external inducer.
  • In the absence of the inducer, HT2-19 cells exhibited significant DNA rereplication.
  • Apoptosis was also observed in HT2-19 cells lacking the inducer, suggesting a role for CDC2 in cell death pathways.

Conclusions:

  • The study successfully established a method for genetically analyzing essential genes in mammalian cells.
  • The findings indicate that human CDC2 is involved in a novel control mechanism preventing premature S-phase entry.
  • This research highlights a previously undetected checkpoint in mammalian cell cycle regulation.

More Related Videos

Zinc-finger Nuclease Enhanced Gene Targeting in Human Embryonic Stem Cells12:13

Zinc-finger Nuclease Enhanced Gene Targeting in Human Embryonic Stem Cells

Published on: August 23, 2014

Non-Viral Engineering of Primary Human T Cells via Homology-Mediated End-Joining Targeted Integration of Large DNA Templates06:10

Non-Viral Engineering of Primary Human T Cells via Homology-Mediated End-Joining Targeted Integration of Large DNA Templates

Published on: May 9, 2025

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.
DNA Damage can Stall the Cell Cycle02:36

DNA Damage can Stall the Cell Cycle

In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...