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CRISPR/Cas Fusions for Imaging

Set Staphylococcus aureus

pCAS9-TP-Sa-dCAS9-eGFP/mRuby2

  • dCas9-3x eGFP/mRuby2 expression vector (dCas9-3x eGFP/mRuby2 driven by the constitutive Ubi4-2 promotor from parsley)
  • MluI site for subcloning of sgRNA
  • Bacterial selection: Spectinomycin (100 mg/l)
  • Plant selection: DL-Phosphinothricin (PPT, 6 mg/l)

 

pEn-Sa-Chimera (sgRNA)

  • sgRNA coding vector (sgRNA can easily be customized for any target site of interest; sgRNA is driven by the Arabidopsis U6-26 promotor
  • sgRNA is not subcloned into pCAS9-TP-Sa-dCAS9-eGFP/mRuby2 via Gateway cloning but via MluI

 

Set Streptococcus pyogenes

pCAS9-TP-Sp-dCAS9-eGFP/mRuby2

  • dCas9-3x eGFP/mRuby2 expression vector (dCas9-3x eGFP/mRuby2 driven by the constitutive Ubi4-2 promotor from parsley)
  • AvrII site for subcloning of sgRNA
  • Bacterial selection: Spectinomycin (100 mg/l)
  • Plant selection: DL-Phosphinothricin (PPT, 6 mg/l)

 

pChimera (sgRNA)

  • sgRNA coding vector (sgRNA can easily be customized for any target site of interest; sgRNA is driven by the Arabidopsis U6-26 promotor
  • sgRNA is not subcloned into pCAS9-TP-Sp-dCAS9-eGFP/mRuby2 via Gateway cloning but via AvrII