B6.129S4(C)-Gt(ROSA)26Sor^{tm1.1(CAG-TMEM192)Dmsa}/J

Stock number: 040002
Product name: LysoTag constitutive, Rosa26-CAG-Tmem192-3xHA
Research areas: Neurobiology Research, Research Tools

Description

LysoTag constitutive (Rosa26-CAG-Tmem192-3xHA) knock-in mice, derived from the conditional LysoTag mutant strain (Stock No. 035401), demonstrate widespread constitutive expression of human TMEM192 cDNA fused to a hemagglutinin (3xHA) tag. The HA tag enables rapid, tissue-specific, anti-HA antibody-driven immunoisolation of intact lysosomes for content profiling.

Detailed description

Mutations in genes encoding lysosomal proteins cause severe disorders collectively known as lysosomal storage diseases (LSDs). Many LSDs are associated with neurodegeneration, indicating a critical role for the lysosome in neuronal homeostasis. Consistent with this, human genetic studies have implicated impaired lysosomal function in several age-associated neurodegenerative diseases, including Alzheimer's disease (AD), Parkinson's disease (PD), and frontotemporal dementia (FTD).

LysoTag constitutive (Rosa26-CAG-Tmem192-3xHA) knock-in mice, derived from the conditional LysoTag mutant strain (Stock No. 035401), constitutively express human TMEM192 cDNA (encoding a lysosomal transmembrane protein) fused to a hemagglutinin (3xHA) tag. Expression of the HA tag enables rapid, tissue-specific, anti-HA antibody-driven immunoisolation of intact lysosomes for content profiling.

Development

A CAG-loxP-neomycin-Stop-loxP-FRT-TMEM192-3xHA-FRT-pA cassette incorporating full-length human TMEM192 cDNA coding sequences fused to a 3x hemagglutinin tag was introduced to intron 1 of the mouse Gt(ROSA)26Sor gene via homologous recombination in a 129S4/SvJae-derived embryonic stem (ES) cells. Resultant conditional LysoTag mice (Stock No. 035401) were backcrossed to C57BL/6J for more than 5 generations by the donating laboratory prior to their arrival at The Jackson Laboratory. Animals from our colony were crossed to CMV-Cre mice (Stock No. 006054) expressing germline cre recombinase from the human cytomegalovirus minimal promoter to excise the floxed Stop and create this LysoTag constitutive allele (Stock No. 040002). Cre recombinase was bred out of the line and sperm was cryopreserved. To establish a live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

Allele

Symbol: Gt(ROSA)26Sor^{tm1.1(CAG-TMEM192)Dmsa}
Name: targeted mutation 1, David Sabatini
MGI accession ID: MGI:8401758
Generation method: Targeted
Attributes: Inserted expressed sequence; Humanized sequence; Epitope tag
Marker symbol: Gt(ROSA)26Sor
Marker name: gene trap ROSA 26, Philippe Soriano
Marker synonyms: beta geo; Gtrgeo26; Gtrosa26; R26; ROSA26; SETD5-AS1; Thumpd3as1
Chromosome: 6
Strain of origin: 129S4/SvJae
Molecular note: The allele Gt(ROSA)26Sor^{tm1.1(CAG-TMEM192)Dmsa} was generated by a CAG-loxP-neomycin-Stop-loxP-FRT-TMEM192-3xHA-FRT-pA cassette incorporating full-length human TMEM192 cDNA coding sequence fused to a 3x hemagglutinin tag introduced into intron 1. Cre recombinase removed the loxP-flanked Stop cassette.