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

Paul Hasty, Ramiro Ramírez-Solis, Robb Krumlauf, Allan Bradley

Research output: Contribution to journalArticle

268 Scopus citations

Abstract

GENE targeting in embryonic stem (ES) cells is a powerful tool for generating mice with null alleles1. Current methods of gene inactivation in ES cells introduce a neomycin gene (neo) cassette both as a mutagen and a selection marker for transfected cells2-11. 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 genes12-16. Inactivation of genes 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).

Original languageEnglish (US)
Pages (from-to)243-246
Number of pages4
JournalNature
Volume350
Issue number6315
DOIs
StatePublished - 1991
Externally publishedYes

ASJC Scopus subject areas

  • General

Fingerprint Dive into the research topics of 'Introduction of a subtle mutation into the Hox-2.6 locus in embryonic stem cells'. Together they form a unique fingerprint.

  • Cite this