Extended Data Fig. 2: Modified sgRNAs enable high frequencies of somatic and heritable editing at the AG locus.

a, Editing efficiency of vectors targeting AG at the eighth systemic leaf. sgRNAs modified with FT (n = 3 independent biological replicates, \(\bar x\) = 0.73 ± 0.19), mFT (n = 3, \(\bar x\) = 0.81 ± 0.09), or Truncated FT (n = 2, \(\bar x\) = 0.85 ± 0.03) yield higher average editing frequencies and lower variance compared to unmodified sgRNAs (n = 4, \(\bar x\) = 0.29 ± 0.29). Tissue was collected approximately 6 weeks after infiltration. The Y axis is the percentage of NGS reads with indels divided by the total number of reads. Each dot represents one plant replicate. Bars indicate the mean editing frequency; error bars represent ± standard deviation. b, sgRNAs modified with FT (n = 3 independent biological replicates, \(\bar x\) = 0.89 ± 0.10, p-value= 0.002) or mFT (n = 3, \(\bar x\) = 0.87 ± 0.22, p-value= 0.011) or Truncated FT (n = 2, \(\bar x\) = 0.81 ± 0.06, p-value= 0.004) produce significantly higher frequencies of heritable editing compared to unmodified sgRNAs (n = 3, \(\bar x\) = 0.10 ± 0.13). In some cases, 100% of progeny contained a mutation. Heritable editing frequency is the fraction, displayed as a percentage, of progeny that contained an indel in at least one allele divided by the total number of progeny genotyped. Each dot represents the heritable editing frequency in progeny of one parent replicate infected with TRV expressing either unmodified sgRNAs (n = 13, 16, or 17) or modified with FT (n = 20, 11, or 14 progeny), mFT (n = 18, 21, or 19) or Truncated FT (n = 20 or 17). Bars indicate the mean editing frequency; error bars represent ± standard deviation. Asterisks indicate statistically significant differences compared to the unmodified sgRNA by the two-sided Student’s t-test, *<0.05, **<0.01. (c) Modified sgRNAs produce a higher portion of bi-allelic and heterozygous mutations compared to unmodified sgRNAs. In one case, 100% of progeny contained bi-allelic mutations. The percentage of each zygosity was determined by the fraction of progeny that contained that genotype divided by total progeny assessed. The same progeny used to determine heritable editing frequency were used to calculate the genotype percentages.