Fig. 1

Schematic diagram of Cas9/sgRNA expression cassette and T-DNA region of the genome editing construct. A Cas9/sgRNA expression cassette contains a 35S promoter driving the expression of Cas9 translationally fused to eGFP, followed by nopaline synthase terminator (Tnos); as well as Arabidopsis U6 promoter (AtU6) driving the expression of sgRNA, followed by AtU6 terminator (TAtU6). The sgRNA is comprised of a scaffold for Cas-binding and 20-nt target sequence. B T-DNA region containing Cas9/sgRNA expression cassette in binary vector pEG301. LB, left border; RB, right border; tMAS, mannopine synthase terminator; Bar, phosphinothricin acetyltransferase conferring BASTA resistance; pMAS, mannopine synthase promoter; HA, human influenza hemagglutinin tag; tOCS, octopine synthase terminator