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Target chromosomes of inducible deletion by a Cre/inverted loxP system in mouse embryonic stem cells

Masako Tada1, Hiroyuki Matsumura, Yuko Kurose

  • 1ReproCELL Inc., Minato-ku, Tokyo, Japan. matada@frontier.kyoto-u.ac.jp

Chromosome Research : an International Journal on the Molecular, Supramolecular and Evolutionary Aspects of Chromosome Biology
|April 1, 2009
PubMed

Insights

Researchers developed a chromosome elimination cassette (CEC) to study chromosomal deletions and their role in cancer. This tool enables the creation of targeted loss of heterozygosity (LOH) events for further genetic mutation studies.

Area of Science:

  • Genetics
  • Cancer Biology
  • Molecular Biology

Background:

  • Chromosomal deletions are implicated in genetic diseases and cancer pathogenesis.
  • These deletions often lead to loss of heterozygosity (LOH) in tumor suppressor genes.
  • Experimental determination of chromosome deletions as cancer triggers is limited by technical challenges.

Purpose of the Study:

  • To develop a novel system for inducing and studying chromosome-wide deletions.
  • To investigate the potential of chromosome deletions as triggers for cancer development.
  • To create a tool for generating Cre-inducible LOH.

Main Methods:

  • Development of a chromosome elimination cassette (CEC) using the Cre/inverted loxP system.
  • Induction of whole or partial chromosome deletions in CEC-transgenic mouse embryonic stem cells (CEC-ESC).
  • Isolation and characterization of CEC-ESC clones with deletions.
  • Analysis of CEC integration sites using fluorescence in-situ hybridization (FISH).

Main Results:

  • Successfully generated diploid CEC-ESC clones with viable small deletions.
  • Identified CEC integration sites on 13 out of 20 mouse chromosome types in 17 analyzed clones.
  • Demonstrated that CEC-tagged chromosomes can be targeted for LOH induction.

Conclusions:

  • The CEC system provides a viable method for studying chromosome deletions.
  • This system facilitates the creation of targeted LOH events for investigating cancer development.
  • The CEC-ESC clones serve as a platform for future genetic mutation studies related to cancer.

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