B6;FVB-Snca^em1Mwcr/J

Stock number: 036763
Product name: Snca^NLS
Research areas: Neurobiology Research

Description

Snca^NLS mice have a nuclear localization signal (NLS) and Flag tag inserted into the 3' region of the endogenous alpha synuclein gene, resulting in localization of aSyn protein to the cell nucleus. These mice exhibit age-dependent motor deficits independent of aSyn aggregation and may be useful in the study of Parkinson's Disease.

Detailed description

The Snca gene encodes the alpha-synuclein (aSyn) protein which is strongly implicated in the pathogenesis of Parkinson's disease.

The Snca^NLS knock-in allele has a nuclear localization sequence and a Flag tag inserted downstream of exon 5 the Snca gene, just before the translational STOP codon. The donating investigator reports visualization of aSyn by immunofluorescence reveals a 1.5-fold increase in the expression in the nuclei of cortical neurons of heterozygotes, and a ~3-fold increase in nuclear aSyn expression in homozygotes despite a ~50% reduction in the total expression of aSyn in brain homogenates. Homozygous Snca^NLS/NLS mice also exhibit age-related deficits in motor behaviors as determined by rotorod and adhesive removal tasks, as well as altered gastric motility and early lethality, with ~25% of homozygotes dying by 20 months of age.
Otherwise, both heterozygotes and homozygotes are viable and fertile with no significant overt phenotypes.

Development

The Snca^NLS allele was generated using CRISPR/Cas9 endonuclease-mediated genome editing in the laboratory of Dr. Maxime Rousseaux (University of Ottawa). A single guide RNA (sgRNA target sequence: TTGGTAGCCTTCCTAATATC) was selected to target the 3' region of the synuclein, alpha gene (Snca) on chromosome 6. A single-stranded oligonucleotide donor DNA template (CACTGTGAAGCAGACAGTTGATATCTGTCACTTCACTGACAAGGCATGCTGTTATTATTTTCTTTTTCTGATATTAGGAAGGCTACCAAGACTATGAGCCTGAAGCCGACTACAAGGACGACGACGACAAGTAAGAATGTCATTGCACCCAATCTCCTAAGATCTGCCGGCTGCTCTTCCATGGCGTACAAGTGCTCAGT) was created encoding a nuclear localization signal (NLS) and Flag tag. These sequences and Cas9 protein were introduced into single cell FVB/N zygotes and transferred to pseudopregnant females. Progeny were screened by DNA sequencing to identify correctly targeted pups, which were then bred to C57BL/6J mice for germline transmission. The resulting Snca^NLS colony was then backcrossed to C57BL/6J mice for 8 generations, and then to C57BL/6NCrl for 2 generations prior to sending males to The Jackson Laboratory Repository in 2021. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6NJ oocytes (Stock No. 005304).

Allele

Symbol: Snca^em1Mwcr
Name: endonuclease-mediated mutation 1, Maxime Rousseaux
MGI accession ID: MGI:6834000
Generation method: Endonuclease-mediated
Attributes: Epitope tag
Synonyms: Snca^NLS
Marker symbol: Snca
Marker name: synuclein, alpha
Marker synonyms: alphaSYN; alpha-Syn; alpha-synuclein; NACP; PARK1; PARK4; PD1
Chromosome: 6
Strain of origin: FVB/N
Molecular note: CRISPR/cas9 genome editing is used to insert a nuclear localization signal (NLS) and FLAG tag. The single-stranded oligonucleotide donor DNA template contains the following sequence: CACTGTGAAGCAGACAGTTGATATCTGTCACTTCACTGACAAGGCATGCTGTTATTATTTTCTTTTTCTGATATTAGGAAGGCTACCAAGACTATGAGCCTGAAGCCGACTACAAGGACGACGACGACAAGTAAGAATGTCATTGCACCCAATCTCCTAAGATCTGCCGGCTGCTCTTCCATGGCGTACAAGTGCTCAGT.