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. 2021 Nov 3;20(2):350–359. doi: 10.1111/pbi.13716

Figure 2.

Figure 2

Targeted mutagenesis induced by CRISPR‐Nm2Cas9 systems in rice. (a) Schematic illustration of the expression cassettes of the CRISPR‐Nm2Cas9 system in the pHUC7b11 binary vectors. The S593Q/W596R double mutations are labelled by a red rectangle inside Nm2Cas9. (b) Mutagenesis efficiencies induced by Nm2Cas9 and Nm2Cas9‐S593Q/W596R in rice. In individual transformants, more than 200 newly emerged calli were collected after 2 weeks of selection under 50 mg/L hygromycin and mixed as a sample for the extraction of genomic DNA. The target region was then analysed by amplicon NGS. The mutagenesis efficiency of Nm2Cas9s was calculated by counting the mutated reads compared with the total clean reads. ±SD values were generated from three biological replicates. *, P < 0.05; **, P < 0.01; two‐tailed t‐test. (c) Identification of targeted mutants in transgenic plants of Nm2Cas9 or Nm2Cas9‐S593Q/W596R.