Transposon-generated ‘knock-out’ and ‘knock-in’ gene-targeting constructs for use in mice

@article{Westphal1997TransposongeneratedA,
  title={Transposon-generated ‘knock-out’ and ‘knock-in’ gene-targeting constructs for use in mice},
  author={Christoph Westphal and Philip Leder},
  journal={Current Biology},
  year={1997},
  volume={7},
  pages={530-533}
}
The conventional technique for targeted mutation of mouse genes entails placing a genomic DNA fragment containing the gene of interest into a vector for fine mapping, followed by cloning of two genomic arms around a selectable neomycin-resistance cassette in a vector containing thymidine kinase [1]; this generally requires 1-2 months of work for each construct. The single 'knock-out' construct is then transfected into mouse embryonic stem (ES) cells, which are subsequently subjected to positive… CONTINUE READING
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