These mutant mice carry a Cre recombinase inducible D620N mutation in exon 15 of the Vps35 (vacuolar protein sorting 35) gene and may be useful in studies of Parkinson's disease.
Dr. Matthew J Farrer, University of British Columbia
Kuldip Dave, The Michael J. Fox Foundation
Vps35 (VPS35 retromer complex component) encodes a component of the heteropentameric retromer complex, which mediates retrograde transport of cargo proteins from endosomes to the trans-Golgi network, endosome to plasma membrane and mitochondria to peroxisomes or lysome. Vsp35 is highly expressed in dopaminergic neurons. The D620N mutation is associated with late-onset autosomal dominant Parkinson’s disease (PD). The mutation alters cargo recognition of substrates and causes alterations in trafficking.
These floxΔneo mice carry a targeted mutation of Vps35 that contains a floxed "mini-cDNA" sequence (exons 15 through 17 and a polyadenylation signal) as well as the D620N mutation in exon 15 that is associated with Parkinson's disease. The polyadenylation signal sequence at the end of the mini-cDNA prevents expression of the D620N mutation. The mice express the wildtype VS35 protein. After Cre recombinase mediated removal of the mini-cDNA, the resulting mice will express the D620N mutation. Homozygotes and heterozygotes are viable and fertile.
When combined with a strain expressing Cre recombinase in the female germline (Stock No. 008454, B6.Cg-Edil3Tg(Sox2-cre)1Amc/J), homozygous offspring exhibit neurodegeneration of DA neurons, accumulation of α-synuclein in DA neurons and age-dependent progressive motor deficits.
Please note: A strain with constitutive expression of D620N is also available, Stock No.023409 B6.Cg-Vps35tm1.1Mjff/J.
A targeted vector containing a floxed "mini-cDNA" (splice acceptor, sequence encoding exons 15 through 17, polyadenylation signal), with a FRT site flanked NEO cassette on the 3'end of the mini-cDNA and a third loxP site, was inserted into intron 14-15. The construct was electroporated into C57BL/6 derived Bruce4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were tested for germline transmission. The mice were then crossed to FLP expressing mice on the C57BL/6J genetic background, to remove the NEO cassette. The FLP recombinase transgene was then bred out from the line. The mice were maintained on the C57BL/6 background. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.
|Allele Name||targeted mutation 1.2,The Michael J Fox Foundation|
|Allele Type||Targeted (Conditional ready (e.g. floxed), No functional change)|
|Allele Synonym(s)||cVKI; VPS35FLOX|
|Gene Symbol and Name||Vps35, VPS35 retromer complex component|
|Strain of Origin||B6.Cg-Thy1a|
|Molecular Note||A targeted vector containing a floxed "mini-cDNA" including a splice acceptor and normal cDNA sequence for exons 15-17, followed by a frt-flanked neo cassette and a third loxP site, inserted into intron 14, and a nucleotide change causing a D620N mutation in the inherent mouse gene sequence 3' of the insertion. The polyadenylation signal sequence at the end of the mini-cDNA prevents expression of the D620N mutation. Flp-mediated recombination removed the neo cassette.|
Heterozygotes and homozygotes are viable and fertile.
When using the floxΔneo Vps35 mouse strain in a publication, please cite the originating article(s) and include JAX stock #021807 in your Materials and Methods section.