High frequency targeting of genes to specific sites in the mammalian genome

Cell. 1986 Feb 14;44(3):419-28. doi: 10.1016/0092-8674(86)90463-0.

Abstract

We corrected a defective gene residing in the chromosome of a mammalian cell by injecting into the nucleus copies of the same gene carrying a different mutation. We determined how the number, the arrangement, and the chromosomal position of the integrated gene, as well as the number of injected molecules influence the gene-targeting frequency. Recombination between the newly introduced DNA and its chromosomal homolog occurred at a frequency of 1 in 10(3) cells receiving DNA. Correction events were mediated by either double reciprocal recombination or gene conversion. This resulted in sequences in the genome being replaced by sequences of the introduced DNA or, in separate experiments, sequences in the incoming DNA being replaced by chromosomal sequences. Both point mutations and deletion mutations were corrected; however, the nature of the mutation carried by the respective sequence influenced whether the integrated or injected sequence was corrected.

MeSH terms

  • Alleles
  • Animals
  • Base Sequence
  • Cell Line
  • Chromosome Deletion
  • DNA Restriction Enzymes
  • DNA, Recombinant
  • Drug Resistance
  • Mice
  • Models, Genetic
  • Mutation*
  • Neomycin / pharmacology
  • Plasmids
  • Recombination, Genetic*
  • Transfection

Substances

  • DNA, Recombinant
  • DNA Restriction Enzymes
  • Neomycin