Fig. 1

Generation of Myt1l mutant mice by CRISPR/Cas9 gene editing. A Schematic of the gene editing strategy. The guide RNA sequence (blue text) targets seven base pairs downstream of the translation start site in exon VI of the mouse Myt1l locus and generates a frameshift deletion. The two forward and one reverse primers (red arrows) used for genotyping are noted. The forward primers, F1 and F2, incorporate the presence and absence of the seven base pairs, respectively. B PCR genotyping of Myt1l mutant mice. As expected, the primer pair F1 and R1 (left lanes) shows a PCR amplification of only wild-type alleles, whereas the primer pair F2 and R1 (right lanes) amplifies only the deletion allele, allowing unequivocal genotyping of wild-type Myt1l^+/+, heterozygous Myt1l^+/-, and homozygous Myt1l^−/− mice. (C) Immunoblotting of Myt1l in WT, HET, and KO E18.5 brains. Whole brain lysates were subjected to Western blotting using two different Myt1l antibodies with similar results. MAP2 was used as a loading control. Note with both antibodies an extra band(*) appeared in mice carrying the mutant allele corresponding to a Myt1l-related protein of ~ 158 kDa