Fig. 1.

Two zebrafish npc1 mutant alleles were generated using CRISPR/Cas9-mediated gene targeting. (A,B) Sequencing chromatographs of npc1^y535 (A), npc1^hg37 (B) and wild-type alleles. Arrowheads delineate the area of the induced insertion/deletion and indicate nucleotide positions corresponding to the wild-type npc1 gene sequence. (C) The npc1^y535 allele disrupts an endogenous AvaII restriction site and genotyping can be performed by restriction fragment length polymorphism analysis. (D) Derived cleaved amplified polymorphic sequence (dCAPS) was used to introduce an AvaII restriction site into PCR products derived from the npc1^hg37 allele that was used for genotyping.