A highly specific SpCas9 variant is identified by in vivo screening in yeast

@inproceedings{Casini2018AHS,
  title={A highly specific SpCas9 variant is identified by in vivo screening in yeast},
  author={Antonio Casini and Michele Olivieri and Gianluca De Petris and Claudia Montagna and Giordano Reginato and Giulia Maule and Francesca Lorenzin and Davide Prandi and Alessandro Romanel and Francesca Demichelis and Alberto Inga and Anna Cereseto},
  booktitle={Nature Biotechnology},
  year={2018}
}
Despite the utility of CRISPR–Cas9 nucleases for genome editing, the potential for off-target activity limits their application, especially for therapeutic purposes. We developed a yeast-based assay to identify optimized Streptococcus pyogenes Cas9 (SpCas9) variants that enables simultaneous evaluation of on- and off-target activity. We screened a library of SpCas9 variants carrying random mutations in the REC3 domain and identified mutations that increased editing accuracy while maintaining… CONTINUE READING

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