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. 2017 Apr 3;7:45763. doi: 10.1038/srep45763

Figure 4. Simultaneous targeted mutagenesis of multiple genes in B. bassiana using the CRISPR/Cas9 system.

Figure 4

(A) Schematic showing homologous replacement of egfp, mp1 and rgs1 by homologous donor DNA-mediated HDR via CRISPR/Cas9 system. (B) PCR screening of the egfp and mp1 double-gene disruptants. The 274 bp and 2.5 kb bands represent the PCR products of the RCPT and the egfp disruptant, respectively. The 701 bp and 2.9 kb bands represent the PCR products of the RCPT and the mp1 disruptant, respectively. (C) PCR screening of the egfp, mp1 and rgs1 triple-gene disruptants. The 274 bp and 2.5 kb bands represent the PCR products of the RCPT and the egfp disruptant, respectively. The 701 bp and 2.9 kb bands represent the PCR products of the RCPT and the mp1 disruptant, respectively. The 638 bp and 2.8 kb bands represent the PCR products of the RCPT and the rgs1 disruptant, respectively. RCPT: recipient strain Bbcas9∆ura5; P: plasmid as positive control; NT: no template control; M: DNA molecular marker.