Fig. 3.
Generation of pcdh1 knockout mice using the CRISPR-Cas9 system. (A) A schematic drawing showing the position and sequences of gRNAs targeting exon2 of pcdh1 gene. Exons (black boxes) and introns (black lines) indicate the structure of one of seven transcription variants of pcdh1 (NM_029357.3). Targeting sequences and PAM sequences of gRNAs are indicated with underline and a gray background, respectively. (B) Genomic sequences around the targeting site of the wild-type (WT) and Δ11 alleles. Targeting sequences of gRNAs are indicated with underline. Dashes: deleted bases. (C) Gross appearance of pcdh1+/+ and pcdh1Δ11/Δ11 female mice at 8 weeks old. (D) Western blotting to detect PCDH1 protein in liver tissue of pcdh1+/+ and pcdh1Δ11/Δ11 adult mice. GAPDH serves as a loading control. (E) Body weight of pcdh1+/+ and pcdh1Δ11/Δ11 mice at 3–8 weeks old. At least three mice were measured for each genotype. Data represent the mean ± SE. A t-test was used for the statistical analyses (*p < 0.05, **p < 0.01, and ***p < 0.001). (F) Body length of pcdh1+/+ and pcdh1Δ11/Δ11 adult male and female mice at 8 weeks old. At least three mice were measured for each genotype. Box plots show the median and 25-75th percentiles (middle line and box, respectively). Whiskers extend to the maximum and minimum data points. A t-test was used for the statistical analyses (*p < 0.05). N.S., not significant