C57BL/6N-Lrrk2^tm1.1Dral/J

Stock number: 039131
Product name: LRRK2 flox-R1441G
Research areas: Neurobiology Research, Research Tools

Description

LRRK2 flox-R1441G mice harbor a Cre-inducible familial Parkinson's disease-associated R1441G missense mutant of the Lrrk2 gene. These mice may be useful for studying Parkinson's disease pathogenesis and neurodegeneration.

Detailed description

Dardarin, also known as leucine-rich repeat kinase 2 (coded by Lrrk2), is involved in regulating inflammation, protein processing, and autophagy. Lrrk2 mutations have been implicated in familial Parkinson's disease. The R1441G clinical mutation occurs in the 'Ras of complex proteins' (ROC) domain of the leucine-rich repeat kinase 2 protein, reducing GTPase activity. These LRRK2 flox-R1441G mice possess loxP sites flanking wildtype exons 31-51 of the Lrrk2 gene and a mutant exon 31, carrying the R1441G missense mutation, inserted immediately downstream of the second loxP site.

Homozygous mice are viable and fertile with no overt phenotype. Prior to Cre-mediated recombination, homozygous LRRK2 flox-R1441G mice demonstrate LRRK2 protein levels in brain tissue similar to wildtype controls. After Cre-mediated recombination, the wildtype exons 31-51 will be excised and the mutant exon 31 with the R1441G mutation is expressed in cre-expressing tissues. These mice may be useful for studying hereditary Parkinson's disease.

Development

The LRRK2 flox-R1441G knock-in allele was designed to incorporate a conditional R1441G point mutation into exon 31 of the leucine-rich repeat kinase 2 (Lrrk2) gene based on the NCBI transcript NM_025730.3 (Ensembl transcript ENSMUST00000060642). The targeting vector, which was inserted into the endogenous Lrrk2 intron 30, consists of an initial loxP site, followed by an FRT-flanked neomycin cassette, exons 31-51 Lrrk2 cDNA including the 3' untranslated region (UTR), an hGHpA (2.1 kb of the human growth hormone gene harboring the hGH polyadenylation signal), an F3-flanked puromycin cassette, and a second loxP site. Genomic exon 31 harboring the R1441G mutation is located immediately downstream of the second loxP site. The target construct was electroporated into C57BL/6N-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts. The resulting chimeric mice were bred to FLP-recombinase mice, on a C57BL/6N background, to remove the FRT-flanked neomycin cassette and the F3-flanked puromycin cassette. The donating laboratory reported they maintained this mouse line on a C57BL/6N background prior to sending homozygous males to The Jackson Laboratory in 2025. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6NJ (Stock No. 005304) oocytes.

In 2025, a 48 SNP (single nucleotide polymorphism) panel analysis, with 43 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the homozygous males sent to The Jackson Laboratory. All markers were C57BL/6N allele-type.

In 2025, the homozygous males sent to The Jackson Laboratory were used to freeze sperm. Samples from those males were analyzed via the miniMUGA 10,000 SNP panel. This showed 98.6-100% of the markers as C57BL/6. The scoring algorithm was unable to discern the C57BL/6 substrain with any certainty. Additionally, one mouse was identified potentially having markers other than C57BL/6 present on Chromosome 19 ~45.45-46.8 Mb (this was not examined further). These data suggest the mice sent to The Jackson Laboratory were on a C57BL/6 genetic background.

Allele

Symbol: Lrrk2^tm1.1Dral
Name: targeted mutation 1.1, Dario R Alessi
MGI accession ID: MGI:7865614
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed)
Marker symbol: Lrrk2
Marker name: leucine-rich repeat kinase 2
Marker synonyms: 4921513O20Rik; 9330188B09Rik; AURA17; cI-46; D630001M17Rik; DARDARIN; LOC381026; PARK8; RIPK7; ROCO2
Chromosome: 15
Strain of origin: C57BL/6N
Molecular note: The allele was designed to incorporate a conditional R1441G point mutation into exon 31 of the leucine-rich repeat kinase 2 (Lrrk2) gene. The targeting vector, which was inserted into the endogenous Lrrk2 intron 30, consists of an initial loxP site, followed by an FRT-flanked neomycin cassette, exons31-51 Lrrk2 cDNA including the 3' untranslated region (UTR), an hGHpA (2.1 kb of the human growth hormone gene harboring the hGH polyadenylation signal), an F3-flanked puromycin cassette, and a second loxP site. Genomic exon 31 harboring the R1441G mutation is located immediately downstream of the second loxP site. FLP-recombinase removed the FRT-flanked neomycin cassette and the F3-flanked puromycin cassette.