Fig. 2
From: CRISPR-FRT targets shared sites in a knock-out collection for off-the-shelf genome editing
Extended applications of CRISPR-FRT. a Members of a plasmid-encoded mutagenesis library (1–4…n) or PCR-amplified linear fragments containing multiple mutated versions of one gene can serve as rescue DNA and thereby be transferred to the chromosome. This allows for single-step transfer of a mutagenesis library to the genome or reconstruction of multiple mutations (red X) in the same gene by only using a single pair of PCR primers to generate the different rescuing templates that contain the desired mutations. b Mutations in essential genes can be delivered to Keio strains with KanR cassettes inserted into the closest neighboring non-essential gene if recombination (dashed lines) occurs beyond the mutation
