C57BL/6NCrl-Aspaem1(G274R)Ccpcz
| Gene Symbol | Aspa |
| Gene Name | aspartoacylase |
| Gene Synonym(s) | ACY2, ASP |
| ENSEMBL | human: ENSG00000108381 mouse: ENSMUSG00000020774 |
| Allele Symbol | C57BL/6NCrl-Aspaem1(G274R)Ccpcz |
| Allele Name | endonuclease-mediated allele 1, Ccpcz |
| Allele Type | point mutation (c.820 G>A, G274R) |
| Genomic location | human: Chromosome 17: 3,472,374-3,503,405 forward strand, GRCh38:CM000679.2 mouse: Chromosome 11: 73,195,818-73,220,422 reverse strand, GRCm39:CM001004.3 |
| Strain of Origin | C57Bl/6NCrl |
| Molecular Note | Mutation introduced by CRISPR/Cas9 double strand break and electroporation of ssODN template with >40 bp homologous arms. Genotyped by PCR amplification of the sequence surrounding the mutation site and subsequent restriction digest by TatI/BsrGI as well as Sanger sequencing. Amino acid sequence is conserved between mice and humans, and the introduced mutation corresponds to the patient mutation. |
| Mutations Made by | Czech Centre of Phenogenomics, Institute of Molecular Genetics of the Czech Academy of Sciences |
Description
Aspartoacylase (EC 3.5.1.15), also called aminoacylase-2, is an enzyme that hydrolyzes N-acetyl-L-aspartic acid (NAA) to aspartate and acetate. Aminoacylase-1 (ACY1; 104620) cleaves acylated L-amino acids, except L-aspartate, into L-amino acids and an acyl group.
Associated pathology
CANAVAN DISEASE
ASPA, c.820 G>A, G274R
Canavan disease is a severe progressive autosomal recessive neurodegenerative disorder characterized by vacuolar degeneration of the lower layers of the brain cortex and the subcortical white matter. Clinical signs usually begin during the first few months after birth, manifested by poor head control and marked developmental delay, and progress to macrocephaly, optic atrophy, seizures, and hypertonia, with death in early childhood. Although most patients with Canavan disease are of Ashkenazi Jewish ancestry, this disorder has also been found in many other ethnic groups (summary by Zeng et al., 2002).
Model Development
We used a synthetic ssODN carrying the target c.820 G>A (G274R) mutation plus one extra silent mutation (ACC>ACA) to create a restriction site for TatI/BsrGI. ssODN had homologous regions on both sides of the mutation >40 bp long. Used ssODN was premixed with Cas9 protein and gRNA and electroporated into zygotes. Zygotes or 2-cell stage embryos were transferred into pseudopregnant recipient mice. Resulting mice (G0 generation) were genotyped by PCR and subsequent restriction digest. The presence of targeted G274R mutation, one extra silent mutation, and no other mutations in the vicinity of targeted region, were confirmed by Sanger sequencing. Positive individuals were backcrossed with C57BL/6NCrl for two generations to establish the colony.