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. 2023 Apr 19;14:1138666. doi: 10.3389/fphar.2023.1138666

FIGURE 6.

FIGURE 6

Generation of MAP6 (STOP) knockout (KO) mice with increased wakefulness and fragmented sleep and wake phases. (A) Schematic of CRISPR/Cas9-engineered MAP6 gene knockout (KO) using C57BL/6 mice as fertilized egg donors. The CRISPR target including the PAM sequence (total 23 bp) contains part of exon 1 of the MAP6 gene that is located 25 nucleotides after the translation start site. ssODN, single-stranded oligodeoxynucleotide. (B,C) Histologic verification of MAP6 KO in the cerebellum of MAP6 (STOP) KO and WT mice. Immunostained MAP6 signals were only detected in wild-type (WT) mice (B) and not in MAP6 (STOP) KO mice (C). Scale bar, 200 μm (D–O) Baseline time course (D–F), total amount (G–I), mean duration (J–L), and episode number (M–O) of SWS, REMS, and wakefulness during the 12h light and 12h dark (shaded areas) phases in MAP6 (STOP) KO and WT mice. (P) Wake episode spectrum in MAP6 (STOP) KO and WT mice. (Q) Mean number of transitions between SWS, wake, and REMS. Data (n = 6/group) are presented as mean ± SEM. Significance levels are indicated as *p < 0.05, **p < 0.01, and ***p < 0.001.