LRRK2 KO2 mice have with exons 29-30 of the Lrrk2 gene deleted. These mice may be useful in studying the cellular function of LRRK2 during aging in the maintenance of protein homeostasis and, specifically, α-synuclein through the regulation of protein degradation pathways.
Jie Shen, Harvard Med Sch/Brigham Women's Hosp
Mice homozygous for the LRRK2 KO2 mutation are viable and fertile, with exons 29-30 (encoding the first half of the Ras-like small GTPase domain) of the Lrrk2 (leucine-rich repeat kinase 2) gene deleted. No mRNA or protein expression from the targeted allele is observed in brain tissue, however some truncated mRNA is observed in kidney tissues. Homozygous mice do not exhibit neurodegeneration or neuropathological changes in the brain. In the kidneys, a tissue where LRRK2 is normally expressed at ~6-fold greater levels than brain, homozygous loss of LRRK2 results in renal atrophy by 20 months of age. This is accompanied by significant (~60-fold) age-dependent accumulation of aggregated α-synuclein and ubiquitinated proteins in the kidney. Specifically, homozygous KO kidneys show widely distributed cytosolic α-synuclein-immunoreactive granular aggregates (some of them may also contain phospho-Ser-129 α-synuclein) or inclusions in boxy cells of renal tubules in the cortical area by 20 months of age. Other kidney defects observed in homozygous mice include impaired ubiquitin-proteasome system (UPS)-mediated protein degradation, impaired autophagy-lysosomal pathway, increased apoptotic cell death, inflammatory response, and oxidative damage. These LRRK2 KO2 mice may be useful in studying the cellular function of LRRK2 during aging in the maintenance of protein homeostasis.
A targeting vector was designed to replace exons 29-30 (which encode the first half of the Ras-like small GTPase domain) of the Lrrk2 (leucine-rich repeat kinase 2) gene with a loxP-flanked PGK-neo cassette. This construct was electroporated into (C57BL/6 x 129)F1-derived MKV6.5 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts. Chimeric mice were bred with B6/129 F1 mice to generate the mutant colony. Next, the mutant mice were bred with CaMKII-Cre transgenic mice on a B6 background. Offspring with the floxed PGK-neo cassette deleted in the germline were bred together (and the CaMKII-Cre transgene was selectively removed) for many generations. Upon arrival at The Jackson Laboratory Repository, mice were bred with B6129SF1/J hybrid mice (Stock No. 101043) for at least one generation to establish the colony.
|Allele Name||targeted mutation 3.1, Jie Shen|
|Allele Type||Targeted (Null/Knockout)|
|Gene Symbol and Name||Lrrk2, leucine-rich repeat kinase 2|
|Strain of Origin||(C57BL/6 x 129)F1|
|Molecular Note||Exons 29 and 30 were replaced with a floxed neo cassette. Cre mediated recombination removed the neo cassette. The absence of protein expression was confirmed by western blot analysis on brain extracts.|
|Mutations Made By|| |
Jie Shen, Harvard Med Sch/Brigham Women's Hosp
When maintaining a live colony, homozygous mice may be bred together.
When using the B6;129-Lrrk2tm3.1Shn/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #016210 in your Materials and Methods section.
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