Fig. 1

通用电气neration ofaitr23456quintuple andaitr123456sextuple mutants.aAlignment of the CRISPR/Cas9 target sequences ofAITR3andAITR4in the Col wild type and theaitr23456quintuple andaitr123456sextuple mutants. The mutants were generated by transforming theaitr256triple andaitr1256quintuple T-DNA insertion mutant plants, respectively with apHEECRISPR/Cas9 construct that simultaneously targetingAITR3andAITR4. Genome editing status in the selected T1 transgenic plants was examined by sequencing. Homozygous Cas9-free mutants were isolated from T2 or T3 progeny of the individual genome edited T1 plants. Numbers indicate the nucleotide position relative to the start condons ofAITR3andAITR4, respectively. Underlines indicate the PAM sites immediately after the target sites.bAmino acid sequence alignment of AITR3 and AITR4 in the Col wild type and theaitr23456quintuple andaitr123456sextuple mutants. The coding sequences ofAITR3andAITR4in theaitr23456quintuple andaitr123456sextuple mutants were used for ORF analysis on ORFfinder (https://www.ncbi.nlm.nih.gov/orffinder/). Predicated amino acid sequences were used for alignment with wild type AITR3 and AITR4, respectively. Numbers indicate amino acid position relative to the first Met residue. Underlines indicate the fully or partial conserved LxLxL motif in AITRs