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. 2020 Jan 9;10:1663. doi: 10.3389/fpls.2019.01663

Figure 1.

Figure 1

The ospin5b/gs3/osmyb30-4 and ospin5b/gs3/osmyb30-25 were precisely edited with CRISPR/Cas9 system. (A) Schematic diagram of the CRISPR/Cas9 vector. The pYLCRISPR/Cas9 binary vector was based on the pCAMBIA1300 backbone which contained the Kanamycin resistance gene. HPT encoded hygromycin B phosphotransferase, which could be driven by the cauliflower mosaic virus 35S promoter (P35S). The Cas9 was driven by the maize ubiquitin promoter (Pubi) and used to edit target sites. Tnos was the terminator of nopaline synthase gene which was chose to terminate the expression of Cas9 gene. U6a, small nuclear RNA promoters, was employed to facilitate the expression of multiple sgRNA cassettes. The six sgRNA cassettes also were inserted behind U6a. (B) Six target sequences alignment in ospin5b/gs3/osmyb30-4 and ospin5b/gs3/osmyb30-25. Blue represents the target sequences. Red represents the changing base.