Figure 2. Generation of alpha kinase 2-knockout (ALPK2-KO) mice.

(a) Sequence of CRISPR RNA (crRNA, red) to target the Cas9 nuclease to a region of exon 3 in the ALPK2 mouse gene. The protospacer adjacent motif (PAM) is shown in green. (b) CRISPR/Cas9-mediated genomic modification was revealed by the T7 endonuclease 1 (T7E1) assay. Six of 16 founder mice contained cleavage products. M, molecular marker; WT, wildtype. (c) Subsequent sub-cloning of flanking regions surrounding the crRNA targeting site (n = 6 sub-clones/mouse) identified three mice (#4, #13, and #15) with frameshift mutations. In mouse #15, the insertion nucleotide is underlined in blue.