Introduction of a subtle mutation into the Hox-2.6 locus in embryonic stem cells

Nature. 1991 Mar 21;350(6315):243-6. doi: 10.1038/350243a0.


Gene targeting in embryonic stem (ES) cells is a powerful tool for generating mice with null alleles. Current methods of gene inactivation in ES cells introduce a neomycin gene (neo) cassette both as a mutagen and a selection marker for transfected cells. Although null alleles are valuable, changes at the nucleotide level of a gene are very important for functional analysis. One gene family in which subtle mutations would be particularly valuable are the clusters of Hox homeobox genes. Inactivation of gene in a cluster with a neo cassette that includes promoter/enhancer elements may deregulate transcription of neighbouring genes and generate a phenotype which is difficult to interpret. We describe here a highly efficient gene targeting method, termed the 'hit and run' procedure. This generates ES cells with subtle site-specific mutations with no selectable marker and may be useful for most genes. We have developed this procedure at the hypoxanthine phosphoribosyltransferase (hprt) locus and subsequently isolated ES cells with a premature stop codon in the homeobox of Hox-2.6 (ref. 14).

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Alleles
  • Animals
  • Base Sequence
  • Chromosome Mapping
  • Electrophoresis, Polyacrylamide Gel
  • Embryo, Mammalian
  • Enhancer Elements, Genetic
  • Genes, Homeobox*
  • Hypoxanthine Phosphoribosyltransferase / genetics
  • Mice
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Promoter Regions, Genetic
  • Recombination, Genetic
  • Stem Cells / metabolism*
  • Transcription, Genetic


  • Hypoxanthine Phosphoribosyltransferase