Figure 1
From: Precise genome-wide base editing by the CRISPR Nickase system in yeast
Limitations of the CRISPR/Cas9 system. (a,b) Schemes of genome editing at an outside area of the PAM and gRNA-targeting sequences by the CRISPR/Cas9 system (a) and by the CRISPR Nickase system (b). (c) Genome editing efficiencies at the CAN1 gene by the conventional method11. The targeted position is described above the graph. (d) Sequencing of the edited CAN1 gene. Cas9 precisely edited within the PAM and gRNA-targeting sequences, but not at a site downstream (DS) from the cleavage site. Introduced stop codons (red) and unintended mutations (inverted) are indicated. The numbers of observed sequences over the numbers of total sequenced strains (e.g., 4/4) are shown. (e) Colony forming efficiencies. The colony forming units (CFUs) on selective medium divided by the CFU of competent cells counted on non-selective medium are presented to evaluate the toxicity of the CRISPR systems. The error bars show standard error of the mean (SEM) based on three independent measurements.
