Figure 2.

Mouse alleles of Copb2. (A) Schematic of the mouse COPB2 protein domains. Asterisks denote sites of mutations. (B–E) The Copb2^Zfn allele is a 5 bp deletion in exon 12 (B, D), which results in a frameshift and creates a premature stop codon (C). (D) Sanger sequencing of the PCR products from a heterozygous animal. Sanger peaks that differ from wild-type are indicated by red arrows and this analysis identifies the precise nature of the deletion. (E) Genotyping was performed by PCR amplification of a 79 bp region surrounding the deletion and gel electrophoresis (4% metaphor agarose). (F–H) The Copb2^R254C (F, G) and Copb2^null (F, H) alleles were created using CRISPR-Cas9 technology targeting exon 8. Red letters indicate nucleotide changes affecting the amino acid sequence while blue letters indicate silent mutations. (I) Copb2^R254C creates an amino acid change orthologous to the patient mutation at amino acid position 254, while Copb2^null creates a frameshift resulting in a premature stop codon (*) at position 264.