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. 2022 Jan 18;11(3):315. doi: 10.3390/cells11030315

Figure 1.

Figure 1

Schematic representation of the binary vector GFP-p1380N-ttLbCas12a:LOBP used to modify Type II LOBP. (a) Pummelo Type II LOBP. Part of the Type II LOBP sequence and its chromatogram were presented, in which EBEPthA4 was highlighted by red rectangles. A crRNA was designed to target EBEPthA4-LOBP, which was indicated by blue. (b) Schematic diagram of GFP-p1380N-ttLbCas12a:LOBP. LB and RB, the left and right borders of the T-DNA region; CsVMV, the cassava vein mosaic virus promoter; GFP, green fluorescent protein; 35T, the cauliflower mosaic virus 35S terminator; CmYLCV, the cestrum yellow leaf curling virus promoter; NosP and NosT, the nopaline synthase gene promoter and its terminator; ttLbCas12a, temperature-tolerant LbCas12a containing the single mutation D156R; AtU6-26, Arabidopsis U6-26 promoter; target, the 23 nucleotides of Type II LOBP highlighted by blue, was located downstream of protospacer-adjacent motif (PAM); HH, the coding sequence of hammerhead ribozyme; HDV, the coding sequence of hepatitis delta virus ribozyme; NptII, the coding sequence of neomycin phosphotransferase II.