C57BL/6-Clptm1^{tm1a(KOMP)Wtsi}/AmcrMmjax

Stock number: 040465
Product name: Clptm1^{tm1a(KOMP)Wtsi} KO-first
Research areas: Cell Biology Research, Developmental Biology Research, Neurobiology Research, Research Tools

Description

Clptm1^{tm1a(KOMP)Wtsi} KO-first mice carry the knock-out first/lacZ reporter allele of the Clptm1 gene. This strain may be useful when studying GABA_A receptor trafficking and inhibitory synaptic transmission.

Detailed description

CLPTM1 (cleft lip and palate associated transmembrane protein 1) is a transmembrane protein involved with regulating GABAergic transmission and modulates inhibitory synaptic strength. Clptm1^{tm1a(KOMP)Wtsi} knock-out (KO)-first mice contain a lacZ/neomycin cassette and a loxP-flanked exon 6. These mice can be converted to a Clptm1 conditional knock-out line by crossing to a FLP-recombinase.

Heterozygous mice are viable and fertile; homozygous mice are not viable. Clptm1^{tm1a(KOMP)Wtsi} KO-first heterozygous mice have elevated surface synaptic GABA_A receptors as well as elevated phasic and tonic inhibitory neurotransmission. This is associated with impaired long-term potentiation and contextual fear memory. These mice may be useful when studying GABA_A receptor trafficking and inhibitory synaptic transmission.

Development

C57BL/6N-derived JM8A3.N1 embryonic stem (ES) cells bearing the Clptm1^{tm1a(KOMP)Wtsi} targeted allele were obtained from Knock-out Mouse Project (KOMP). The targeting cassette is composed of an FRT site followed by lacZ sequence, an initial loxP site, a neomycin resistance sequence, a second FRT site, and a second loxP site upstream of exon 6 of the cleft lip and palate associated transmembrane protein 1 (Clptm1) gene. A third loxP site is inserted downstream of exon 6. Targeted ES cells were injected into recipient blastocysts. The resulting chimeric mice were bred to C57BL/6J to confirm germline transmission. The Clptm1^{tm1a(KOMP)Wtsi} knock-out (KO)-first mouse line was then backcrossed to C57BL/6J for 11 generations by the donating laboratory. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

Allele

Symbol: Clptm1^{tm1a(KOMP)Wtsi}
Name: targeted mutation 1a, Wellcome Trust Sanger Institute
MGI accession ID: MGI:4419487
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Reporter; Null/Knockout
Synonyms: Clptm1^{tm1a(KOMP)Wtsi} "KO-first"; KOfirst
Marker symbol: Clptm1
Marker name: cleft lip and palate associated transmembrane protein 1
Marker synonyms: HS9; N14
Chromosome: 7
Strain of origin: C57BL/6N-A^tm1Brd
Molecular note: The L1L2_Bact_P cassette was inserted at position 19372598 of Chromosome 7 upstream of the critical exon 6 (Build GRCm39). The cassette is composed of an FRT site followed by lacZ sequence and a loxP site. This first loxP site is followed by a neomycin resistance gene under the control of the human beta-actin promoter, SV40 polyA, a second FRT site and a second loxP site. A third loxP site is inserted downstream of exon 6 at position 19373363. Exon 6 is thus flanked by loxP sites. A "conditional ready" (floxed) allele can be created by flp recombinase expression in mice carrying this allele. Subsequent cre expression results in a knockout mouse. If cre expression occurs without flp expression, a reporter knockout mouse will be created. Western blot analysis confirmed disruption of protein expression in brain tissue lysates from E18 homozygous embryos. Further information on targeting strategies used for this and other IKMC alleles can be found at http://www.informatics.jax.org/mgihome/nomen/IKMC_schematics.shtml
General note: Cell line EPD0395_5_D10 was successfully used to make chimeric mice. Germline transmission was accomplished. J:357551.