Fig. 1.

Fig. 1. Multiplex genome editing through Cas12f1-mediated negative selection. Mutations are introduced into the galK and xylB genes by oligonucleotides. sgRNAs are expressed from a dual sgRNA plasmid and form a complex with Cas12f1 nuclease. The scaffold sequences of both sgRNAs are identical and indicated by gray lines. Each sgRNA has a targeting region sequence (TRS) specific to either galK or xylB. Unedited targets are cleaved by the sgRNA/Cas12f1 complex. However, cells with edited targets are not cleaved, facilitating the identification of edited cells through negative selection.

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