Conventional gene targeting protocols lead to loss of targeted cells when applied to a silent gene locus in primary fibroblasts.

@article{Marques2006ConventionalGT,
  title={Conventional gene targeting protocols lead to loss of targeted cells when applied to a silent gene locus in primary fibroblasts.},
  author={Margarita M Marques and Alison J. Thomson and Kenneth J. McCreath and Jim McWhir},
  journal={Journal of biotechnology},
  year={2006},
  volume={125 2},
  pages={185-93}
}
Gene targeting in livestock fibroblasts has proven difficult to achieve, particularly if the target gene is silent. We first tested whether efficient gene targeting at the transcriptionally active ovine alpha1(I) procollagen (COL1A1) locus required the use of a promoter trap vector. We compared gene targeting frequencies at the ovine COL1A1 locus using both a promoter trap and a non-promoter trap selection strategy. We demonstrated that targeted cells could be isolated regardless of whether an… CONTINUE READING