Figure 3
From: A simple method using CRISPR-Cas9 to knock-out genes in murine cancerous cell lines
The blunt-end of the cassette with selection marker and single-strand oligodeoxynucleotides (ssODNs) were critical for increasing the efficiency to knock the cassette into genome. (A, B) The schemas of knockout of the target gene by pX330 plasmids, ssODNs, and un-cut plasmid with selection marker (A) or sticky ends of DNA coding selection marker cut by NotI and AscI (B). (C) The schema of knockout of the target gene by pX330 plasmids and the DNA with selection marker smoothly cut by EcoRV without ssODNs. Agarose gel images showing the genotyping results. The tables indicate the genotyping results of 93 clones. 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.
