Directed evolution of an enhanced and highly efficient FokI cleavage domain for zinc finger nucleases.

@article{Guo2010DirectedEO,
  title={Directed evolution of an enhanced and highly efficient FokI cleavage domain for zinc finger nucleases.},
  author={Jing Guo and Thomas Gaj and Carlos F. Barbas},
  journal={Journal of molecular biology},
  year={2010},
  volume={400 1},
  pages={
          96-107
        }
}
Zinc finger nucleases (ZFNs) are powerful tools for gene therapy and genetic engineering. The high specificity and affinity of these chimeric enzymes are based on custom-designed zinc finger proteins (ZFPs). To improve the performance of existing ZFN technology, we developed an in vivo evolution-based approach to improve the efficacy of the FokI cleavage domain (FCD). After multiple rounds of cycling mutagenesis and DNA shuffling, a more efficient nuclease variant (Sharkey) was generated. In… CONTINUE READING
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