Figure 2 | Scientific Reports

Figure 2

From: High-frequency random DNA insertions upon co-delivery of CRISPR-Cas9 ribonucleoprotein and selectable marker plasmid in rice

Figure 2

Schematic illustration of plasmid constructs used for rice transformation. (A) pCAMBIA1301 (Roberts et al. 1996) used for the co-delivery of CRISPR RNA complex and plasmid DNA. (B) pDW3586 (this work) used for the construction of pTF6005 and pTF6005, which carries PDS gRNA1 and gRNA2, respectively. RB, right border; LB, left border; 2xP35S-hptII-T35S, hygromycin resistance gene cassette; P35S-gus-Tnos, beta-glucuronidase (GUS) report gene cassette; PZmUbi-Cas9-T35S, maize ubiquitine promoter driving Cas9 expression cassette; OsU6, Oryza sativa U6 small RNA promoter; SacB cassette, B. subtilis counter-selectable marker gene for facilitating the cloning of gRNAs; MCS, multiple cloning site; pVS1, replication origin from Pseudomonas aeruginosa; bom, E. coli origin of transfer; colE1, replication origin plasmid ColE1; KanR, kanamycin resistance gene; SpR, spectinomycin resistance gene; numbers on the plasmids refer to sequence coordinates.

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