B6C3-Atp1a3^em3Lutzy/Mmjax

Stock number: 038294
Product name: Atp1a3^{D801N L802L}
Research areas: Developmental Biology Research, Mouse/Human Gene Homologs, Neurobiology Research, Research Tools

Description

Atp1a3^D801N is a mutant of the ATPase, Na+/K+ transporting, alpha 3 polypeptide gene carrying the D801N point mutation. These mice may be useful in studying dystonia-12, alternating hemiplegia of childhood-2, and cerebellar ataxia, areflexia, pes cavus, optic atrophy, and sensorineural hearing loss (CAPOS).

Of note, MMRRC distributes the same Atp1a3^D801N mutation on a C57BL/6J background as RRID:MMRRC_066968-JAX.

Detailed description

CRISPR/cas9 endonuclease mediated genome editing of the ATPase, Na+/K+ transporting, alpha 3 polypeptide gene (Atp1a3) was used to introduce the D801N point mutation. This mutation has been identified in humans with alternating hemiplegia of childhood (AHC). The targeted Atp1a3 gene encodes the alpha 3 catalytic subunit of the sodium/potassium transporting ATPase transmembrane ion pump. Mutations in this gene have been associated with dystonia-12, alternating hemiplegia of childhood-2, and cerebellar ataxia, areflexia, pes cavus, optic atrophy, and sensorineural hearing loss (CAPOS).

On the C57BL/6J background (MMRRC Stock No. 066968), mice submerged in cold water (5-10°C) and/or in a cold environment (-20°C) undergo abrupt hypothermia-induced dystonia (HID). Furthermore, heterozygous mice show severe survival defects with 50% of mice failing to reach wean age and heterozygous mice generally fail to survive past 35 weeks of age likely due to sudden, spontaneous seizures. To improve survival defects, hybrid vigor was introduced by transferring the D801N mutation onto an F1 genetic background with C3H/HeJ. In contrast to C57BL/6 heterozygotes, B6C3 heterozygotes exhibit survival defects with a later onset at ~ 10 weeks of age and ~50% of heterozygous mice reach 35 weeks of age. Nevertheless, heterozygoyes still undergo rapid dystonia when submerged in cold water (5°C) at various, tested ages such as 6, 10 and 13 weeks of age. In addition, motor and motor learning defects (rotarod) were observed/tested as early as 10 weeks of age. Of note, heterozygous females demonstrate a better survival compared to heterozygous males.

Development

Plasmids encoding a signal guide RNA designed to introduce the D801N point mutation in exon 17 of the Atp1a3 gene on chromosome 7 and the cas9 nuclease were introduced into the cytoplasm C57BL/6J-derived fertilized eggs with well recognized pronuclei. An additional silent nucleotide change was introduced as part of the CRISPR/cas9 targeting strategy. This change of CTG to CTA (L802L) has no effect on protein sequence but could impact potential genotyping strategies. Correctly targeted embryos were transferred to pseudopregnant females. Correctly targeted pups were identified by sequencing and PCR and further bred to C57BL/6J to develop the C57BL/6J-Atp1a3^em3Lutzy/Mmjax (MMRRC Stock No. 066968). Sperm from C57BL/6J-Atp1a3^em3Lutzy/Mmjax males was injected into C3H/HeJ females to create this strain. This strain is maintained as a cross of heterozygous Atp1a3 ^D801N/+ mice to B6C3F1/J mice at each generation.

Allele

Symbol: Atp1a3^em3Lutzy
Name: endonuclease-mediated mutation 3, Cathy Lutz
MGI accession ID: MGI:6438218
Generation method: Endonuclease-mediated
Attributes: Not Applicable
Synonyms: Atp1a3^{D801N L802L}
Marker symbol: Atp1a3
Marker name: ATPase, Na+/K+ transporting, alpha 3 polypeptide
Marker synonyms: AHC2; Atpa1a3; Atpa-2; CAPOS; DEE99; DYT12; MGC:27631; MGC:38713; MGC:38914; MGC:39036; RDP
Chromosome: 7
Strain of origin: Not Specified
Molecular note: Plasmids encoding a signal guide RNA are designed to introduce the D801N point mutation in exon 17. An additional silent nucleotide change was introduced as part of the CRISPR/Cas9 targeting strategy. This change of CTG to CTA (L802L) has no effect on protein sequence. Correctly targeted pups were identified by sequencing and PCR.