129S-Rpe65^tm1Lrcb/Mmjax

Stock number: 040271
Product name: Rpe65^R91W
Strain synonyms: Rpe65R91W , Rpe65 R91W , Rpe65-R91W , Rpe65(R91W)
Research areas: Developmental Biology Research, Mouse/Human Gene Homologs, Research Tools, Sensorineural Research

Description

Rpe65^R91W mice express a clinical missense mutation (R91W) and may be useful for studying retinal diseases and potential therapeutics.

Detailed description

Rpe65 encodes a protein expressed specifically in the retinal pigment epithelium (RPE), a monolayer of cells that closely interacts with the photoreceptors of the retina and plays essential roles in the maintenance of visual function. These Rpe65^R91W mice express an arginine to tryptophan missense mutation (R91W) in the Rep65 gene. This mutation results in reduced levels of RPE65 ultimately leading to early-onset retinal dystrophy. Homozygous mice are viable and fertile, but do demonstrate progressive visual impairment starting at 4 weeks of age. Heterozygous mice do not have any visual impairments. Rpe65^R91W mice may be useful for studying inherited retinal dystrophies.

Development

The Rpe65^R91W targeting construct was designed to introduce two point mutations (CGA > TGG) into the retinal pigment epithelium 65 (Rpe65) gene, resulting an arginine to tryptophan amino acid substitution. The targeting vector also contained a loxP-flanked neomycin cassette and a diphtheria toxin (DT) cassette as selection markers. The construct was electroporated into 129S6/SvEvTac-derived TC1 embryonic stem (ES) cells and correctly targeted ES cells were injected into recipient blastocysts. The resulting chimeric mice were mated to 129S6 mice for germline transmission. Rpe65^R91W mice were then crossed to a germline Cre-deleter mouse line (JAX Stock No. 003960) to remove the floxed neomycin cassette. The Rpe65^R91W progeny were maintained on a 129S6 background by the donating laboratory. Upon arrival, sperm was cryopreserved. To establish a live colony, an aliquot of frozen sperm was used to fertilize 129S1/SvImJ oocytes (Stock No. 002448).

Allele

Symbol: Rpe65^tm1Lrcb
Name: targeted mutation 1, Christian Grimm
MGI accession ID: MGI:3771723
Generation method: Targeted
Synonyms: R91W; Rpe65^R91W
Marker symbol: Rpe65
Marker name: retinal pigment epithelium 65
Marker synonyms: A930029L06Rik; BCO3; LCA2; Mord1; mRPE65; p63; rd12; RP20; sRPE65
Chromosome: 3
Strain of origin: 129S6/SvEvTac
Molecular note: Two point mutations were introduced into exon 4 at codon 91 (CGA to TGG) to cause an arginine to tryptophan missense mutation (R91W). A floxed neomycin resistance cassette was also introduced into intron 6 for targeting purposes. Founder mice were mated with deleter mice to remove the neomycin cassette, leaving behind a single loxP site. Levels of the mutant protein are reduced by 95% compared to wild-type controls as determined by western blots of retinal extracts.