Fig. 1

Analysis of CRISPR/Cas9-induced targeted gene mutations in Arabidopsis RNA-silencing pathway mutants. a An ectopically expressed CRISPR/Cas9 system (pIUC-TT4) designed for targeted gene mutagenesis in Arabidopsis. pAtU6, Arabidopsis U6-26 promoter; sgRNA, single guide RNA; pAtUBQ1, Arabidopsis UBIQUITIN 1 promoter; NLS, SV40 nuclear localization signal; SpCas9, Streptococcus pyogenes Cas9 gene; tAtUBQ1, Arabidopsis UBIQUITIN 1 terminator. b Schematic diagram of the TT4 gene. The vertical arrows indicate the positions of the sgRNA target sites. The horizontal arrows indicate the position of primers used for analyzing the relative integrity (RI) of TT4 gene. c–e T1 seeds obtained from pIUC-TT4 transgenic lines exhibited different severities of seed coat phenotypes. c Wild-type seeds. d Chimeric seeds. e Mutated seeds. f Phenotype analysis of CRISRP/Cas9-induced mutations in different silencing pathway mutants. % of mutation = (chimera + mutant)/total