Figure 1.

Targeted mutagenesis induced by the CRISPR‐Nm1Cas9 system in rice. (a) Schematic illustration of the expression cassettes of the CRISPR‐Nm1Cas9 system in the pHUC7a11 binary vector. OsU3, rice U3 promoter; ZmUBI, maize ubiquitin‐1 promoter; NLS, nuclear location sequence; poly T, poly T terminator; 35S ter, CaMV 35S terminator. (b) Schematic representation of the 121‐nt sgRNA (esgRNA) scaffold of Nm1Cas9. The red letters indicate the discrepant nucleotides existing in the 121‐nt scaffold but not in the 101‐nt sgRNA scaffold. (c) Identification of targeted mutants in CRISPR‐Nm1Cas9 transgenic plants. The target sequence is presented above the column. Blue letters indicate the protospacer, and the PAM is labelled in orange. For each vector, 48 regenerated T₀ plants were examined, and the ratio of the number of plants with certain zygotypes to the total plant number was considered the frequency. WT, wild type at the target; He, heterozygous mutant (a mutated allele and a WT allele); Ch, chimeric mutant (more than three different alleles at the target in a single plant); Bi, biallelic mutant (two different mutated alleles); Ho, homozygous mutant (single mutated allele); sgRNA, 101‐nt sgRNA scaffold used in the CRISPR‐Nm1Cas9 system; esgRNA, 121‐nt sgRNA scaffold.