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

Chromosome Res. 2009;17(4):443-50. doi: 10.1007/s10577-009-9035-0. Epub 2009 Mar 31.

Abstract

Chromosomal deletions are widely involved in serious genetic diseases and in the pathogenesis of cancers. These deletions often generate loss of heterozygosity (LOH) of one of the alleles of a tumor suppressor gene. Because of the technical difficulty inherent in genetic manipulation studies of a chromosome-wide deficiency, it has not been experimentally determined whether chromosome deletions could be a trigger for cancer development. Using the Cre/inverted loxP system, we have developed a chromosome elimination cassette (CEC) that Cre-dependently induces whole or partial deletions of the CEC-tagged chromosomes. Most deletions are usually fatal, but diploid cells carrying small deletions have been obtained from mouse embryonic stem cells carrying a CEC transgene (CEC-ESC). Here, we further isolated various CEC-ESC clones and analyzed CEC integration sites using the fluorescence in-situ hybridization method. In 17 CEC-ESC clones possessing normal chromosome sets, 13 types of chromosomes out of 20 pairs of mouse chromosomes were tagged by CEC. Each CEC-tagged chromosome could become a future target for the creation of a Cre-inducible LOH by a combination of in vitro and in vivo genetic mutation.

MeSH terms

  • Animals
  • Cell Line
  • Chromosome Deletion*
  • Chromosomes / metabolism*
  • Clone Cells
  • Embryonic Stem Cells / metabolism*
  • Fluorescein-5-isothiocyanate / metabolism
  • Fluorescent Dyes / metabolism
  • Genes, Reporter
  • Green Fluorescent Proteins / metabolism
  • In Situ Hybridization, Fluorescence
  • Indoles / metabolism
  • Integrases / genetics
  • Integrases / metabolism
  • Loss of Heterozygosity*
  • Metaphase
  • Mice
  • Mice, Transgenic
  • Physical Chromosome Mapping
  • Recombination, Genetic*

Substances

  • Fluorescent Dyes
  • Indoles
  • Green Fluorescent Proteins
  • DAPI
  • Cre recombinase
  • Integrases
  • Fluorescein-5-isothiocyanate