This Park2 (Parkin S65A KI) point mutation knock-in strain constitutively expresses mutant S65A Parkin. These mice may be useful in studies of Parkinson's disease.
Miratul Muqit, University of Dundee
The targeted Park2 gene encodes a protein that is part of the E3 ubiquitin ligase complex. Mutations in this gene have been associated with Parkinson disease. As the mice are further characterized, we will modify the strain description and add phenotype data. These Parkin S65A point mutation knock-in mice constitutively expresses mutant S65A Parkin.
Homozygotes are viable, fertile and do not exhibit any overt phenotype up to 1 year of age. Mutant mice over 1 year in age are currently being assessed (July 2016).
Expression of Park2 is unaltered in brain tissue (additional tissues are being analyzed). As the mice are further characterized, we will modify the strain description and add phenotype data.
A targeting vector containing an FRT site flanked PuroR selection cassette was used to insert the S65A point mutation in exon 3 of the Park2 (Parkin) gene. The FRT site flanked PuroR selection cassette was inserted into intron 2.
The construct was electroporated into the TaconicArtemis C57BL/6NTac derived Art B6 3.6 embryonic stem (ES) cells.
Correctly targeted ES cells were injected into BALB/c blastocysts.
The resulting chimeric male animals were bred to C67BL/6 females to achieve germline transmission.
The mice were crossed to FLP recombinase expressing transgenic mice, C57BL/6-Tg(CAG-Flpe)2Arte, to remove the FRT site flanked selection cassette, and resulting in the constitutive expression of the mutant S65A exon 3.
The mice were then backcrossed to C57BL/6J for 9 generations.
Cryopreserved sperm was transferred to The Jackson Laboratory. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).
|Allele Name||targeted mutation 1.1, Miratul Muqit|
|Allele Type||Targeted (Not Applicable)|
|Allele Synonym(s)||Parkin S65A|
|Gene Symbol and Name||Prkn, parkin RBR E3 ubiquitin protein ligase|
|Strain of Origin||C57BL/6NTac|
|Molecular Note||The targeting vector contains an FRT-flanked PuroR selection cassette inserted into intron 2 and an S65A point mutation inserted into exon 3. Flp-mediated recombination removed the FRT-flanked PuroR cassette.|
When maintaining a live colony, these mice can be bred as homozygotes. Homozygotes are viable, fertile, and do not exhibit any overt phenotype up to 1 year of age. Mutant mice over 1 year in age are currently being assessed (July 2016). As the mice are further characterized, we will modify the strain description and add phenotype data.
When using the Parkin S65A KI mouse strain in a publication, please cite the originating article(s) and include JAX stock #029247 in your Materials and Methods section.