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Efficient CRISPR/Cas9-based genome editing and its application to conditional genetic analysis in Marchantia polymorpha.

Shigeo S SuganoRyuichi NishihamaMakoto ShirakawaJunpei TakagiYoriko MatsudaSakiko IshidaTomoo ShimadaIkuko Hara-NishimuraKeishi OsakabeTakayuki Kohchi
Published in: PloS one (2018)
Marchantia polymorpha is one of the model species of basal land plants. Although CRISPR/Cas9-based genome editing has already been demonstrated for this plant, the efficiency was too low to apply to functional analysis. In this study, we show the establishment of CRISPR/Cas9 genome editing vectors with high efficiency for both construction and genome editing. Codon optimization of Cas9 to Arabidopsis achieved over 70% genome editing efficiency at two loci tested. Systematic assessment revealed that guide sequences of 17 nt or shorter dramatically decreased this efficiency. We also demonstrated that a combinatorial use of this system and a floxed complementation construct enabled conditional analysis of a nearly essential gene. This study reports that simple, rapid, and efficient genome editing is feasible with the series of developed vectors.
Keyphrases
  • genome editing
  • crispr cas
  • genome wide
  • transcription factor
  • emergency department
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  • single cell
  • adverse drug
  • electronic health record
  • data analysis
  • loop mediated isothermal amplification