Figure 11.

ABC-CRISPR of Slp/Sik3 rescues hypersomnia of Sik3^Slp/+;Rosa26^Cas9/+ mice. A, Schematic showing triple sgRNA target sites within exons 3, 4, and 5 of Sik3 gene. B, Quantitative analysis of indel mutations at the sgRNA target sites and 21 predicted off-target sites for the three sgRNAs targeting Sik3 gene. C, Immunoblotting of whole-brain lysates from AAV-sgRNA^NT and AAV-sgRNA^Sik3 injected Sik3^Slp/+; Rosa26^Cas9/+ mice with anti-SIK3 and anti-ACTIN antibodies. D, Quantification of the levels of SIK3-L/SLP-L and SIK3-S/SLP-S proteins shown in C (n = 3). E–G, Hourly plot of NREMS time (E), quantification of total NREMS time (F), and hourly plot of NREMS δ power (G) in the AAV-sgRNA^NT (n = 7) or AAV-sgRNA^Sik3 (n = 7) injected Sik3^Slp/+; Rosa26^Cas9/+ mice. H–L, Hourly plots of Wake (H) or REMS (J) time, quantification of total Wake (I) or REMS (K) time, and EEG power spectra analysis of NREMS, REMS, and Wake states (L) in the AAV-sgRNA^NT (n = 7) or AAV-sgRNA^Sik3 (n = 7) injected Sik3^Slp/+; Rosa26^Cas9/+ mice. Data are mean ± SEM. D, Unpaired t test. E–L, Two-way ANOVA with Dunn's multiple comparisons test. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.