Figure 2
From: A simple method using CRISPR-Cas9 to knock-out genes in murine cancerous cell lines
SUCCESS induced homozygotic deletion of the target gene. (A) The design of the selection markers is shown. Knock-in efficiency of the selection marker were calculated using genotype results of 93 clones. Arrows show 80mer ssODNs consisting of a 40mer homology sequence to the target genomic region and another 40mer homology sequence to the cassette with selection marker. The tables indicate the results of genotyping of 93 clones. BSD, blasticidin S resistant gene; HygR, hygromycin resistant gene; NeoR, neomycin resistant gene; PuroR, puromycin resistant gene. (B) The schema of ligation between the target genomic region and the cassette with selection marker, both at 5′ and 3′ sites. The ligation was analyzed by direct sequencing. 5′, 5′ site between the genomic region and the cassette; 3′, 3′ site between the genomic region and the cassette; WT, wild-type allele; −/−, homozygously knockout clones: −/−*, homozygously knockout clones without the detection of 5′ or 3′ sites ligation between the genomic region and the cassette; %KI, percentage of knock-in in 93 clones.
