CRISPR‐Cas9‐mediated efficient directed mutagenesis and RAD51‐dependent and RAD51‐independent gene targeting in the moss Physcomitrella patens

@inproceedings{Collonnier2017CRISPRCas9mediatedE,
  title={
CRISPR‐Cas9‐mediated efficient directed mutagenesis and RAD51‐dependent and RAD51‐independent gene targeting in the moss Physcomitrella patens
},
  author={C{\'e}cile Collonnier and Aline Epert and Kostlend Mara and François Maclot and Anouchka Guyon‐Debast and Florence Charlot and Charles White and Didier G. Schaefer and Fabien Nogu{\'e}},
  booktitle={Plant biotechnology journal},
  year={2017}
}
The ability to address the CRISPR-Cas9 nuclease complex to any target DNA using customizable single-guide RNAs has now permitted genome engineering in many species. Here, we report its first successful use in a nonvascular plant, the moss Physcomitrella patens. Single-guide RNAs (sgRNAs) were designed to target an endogenous reporter gene, PpAPT, whose inactivation confers resistance to 2-fluoroadenine. Transformation of moss protoplasts with these sgRNAs and the Cas9 coding sequence from… CONTINUE READING
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