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. 2017 Nov 30;7:16624. doi: 10.1038/s41598-017-16998-8

Figure 1.

Figure 1

Schematic of the CAGO genome-editing technique and the structure of the pCAGO plasmid. (A) The editing cassette is inserted into the target locus by homologous recombination. Subsequently, the CRISPR/Cas9 system is expressed to generate a double-strand break (DSB) at the inserted universal N20PAM sequence, after which a DSB-mediated intra-chromosomal recombination event between the L and L_short homologous arms completes the markerless genome editing. (B) Structure of the pCAGO functional region; the λ-Red system is induced using the Ptrc promoter, cas9 is induced using the pBAD promoter, and gRNA is expressed by a constitutive promoter. The temperature-sensitive repA101ts is used as the plasmid replicon for easy curing afterwards.

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