Fig. 1
From: Genome editing through large insertion leads to the skipping of targeted exon
Strategy for ZFN-mediated generation of knockout cell lines. a Exon 9 of hCDC14A was targeted by zinc finger nuclease (ZFN). A donor template containing two homologous arms (HA), stop codon and neomycin selection cassette was used for their homologous recombination mediated insertion within the double strand break (DSB) site. Junction PCR with forward primer (blue arrow) in NeoR cassette and reverse primer (red arrow) in the genome outside homology arm confirmed successful targeting and insertion of the selection marker. b Exon 4 was targeted to knockout hCDC14B gene. As donor template for hCDC14A, neomycin selection cassette was used to generate hCDC14B single knockout. Puromycin cassette was used when hCDC14B was knocked-out on top of hCDC14A −/− cells (neomycin). Forward (blue) and reverse (red) primers used for the junction PCR are shown in the figure