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 language | English (US) |
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Pages (from-to) | 243-246 |
Number of pages | 4 |
Journal | Nature |
Volume | 350 |
Issue number | 6315 |
DOIs | |
State | Published - 1991 |
Externally published | Yes |
ASJC Scopus subject areas
- General