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. 2023 Oct 20;14:1233418. doi: 10.3389/fpls.2023.1233418

Figure 4.

Figure 4

(A) Position of the sgRNAs targeting the raffinose synthase (PvRS1 and PvRS2) and stachyose synthase (PvSS) genes in P. vulgaris. (B) Schematic representation of the T-DNA of the pMR356-GFP and pMR394-GFP vectors. The A. thaliana codon-optimized Cas9 gene (Atco-Cas9) was driven by a Parsley Ubiquitin promoter (PcUbi) in pMR356-GFP whereas Atco-Cas9 was driven by a 2X35S-Ω promoter in pMR394-GFP. In both vectors, the Atco-Cas9 gene was fused to a nuclear localization signal derived from simian virus 40 (SV40 NLS) and transcription was terminated by a A. thaliana heat shock protein 18.2 terminator (AtHSP18.2 ter). The transcription of the sgRNA was driven by a U6 promoter. The enhanced green fluorescent protein (eGFP) gene was placed under the regulation of a Cauliflower Mosaic Virus 35S promoter (CaMV 35S) and nopaline synthase terminator (NOS ter).