Fig. 1

Targeted editing of TMT1 gene in 41B grape cells. A Schematic illustration of target design and LbCas12a expression vector. NPTII, neomycin phosphotransferase gene; NOS, terminator of nopaline synthase gene; OCS, terminator of octopine synthase gene; EGFP, enhanced green fluorescent protein; DR, LbCas12a direct repeat; LB, left border; RB, right border. B Transformed 41B cells with EGFP fluorescence. The LbCas12a vector containing the TMT1 crRNA array (LbCas12a-TMT1) and empty vector (EV) without crRNA were introduced into 41B cells, respectively. C Identification of exogenous T-DNA insertions by PCR. LbCas12a-specific primers were used to identify the T-DNA in 41B cells. The CRISPR vector and wild-type (WT) cells were used as positive (P) and negative controls, respectively. D Mutation efficiencies detected for EV and LbCas12a-TMT1 cells. E Representative mutated sequences detected at the two targets in TMT1 gene. The target sequences and PAMs are indicated in red and blue, respectively. The mutation types are shown on the right. WT, wild-type sequences; Mut, mutated sequences. F Deletion size of nucleotides observed at the two targets in TMT1 gene. G Positions of nucleotide deletions happened at the two targets in TMT1 gene. The first base adjacent to the PAM is referred to as position 1. Data are collected from three replicates and shown as means ± SD