CRISPR/Cas Fusions for Imaging

Set Staphylococcus aureus

pCAS9-TP-Sa-dCAS9-eGFP/mRuby2 (sequence information GFP/mRuby)

• 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) (sequence information)

• 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 (sequence information GFP/mRuby)

• 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) (sequence information)

• 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