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. 2015 Apr 3;1(3):e1500248. doi: 10.1126/sciadv.1500248

Fig. 1. Candida CRISPR expression constructs.

Fig. 1

(A) Duet system consists of two plasmids, pV1025 (top before and after flipout), which targets ENO1, and pV1090, which targets RP10. (B) Solo system consists of one plasmid, pV1093, which targets ENO1. Figures are not drawn to scale. (C) Both Solo and Duet guide expression systems permit rapid cloning by digestion with BsmBI followed by ligation of annealed oligos (shaded sequences) with desired guide sequence (ADE2 guide sequence in red box). (D) Schematic of Cas9 mutagenesis method. The system can create homozygous mutations in the gene (*) and simultaneously mutate sequences (for example, the PAM) to prevent repeated cleavage subsequent to integration.