B6.129(SJL)-Grip1^tm1Rlh/J

Stock number: 024414
Research areas: Neurobiology Research, Research Tools

Description

The first PDZ domain of Grip1 is flanked by loxP sites in this conditional knockout strain. This enables the development of Cre-mediated knockouts useful in studies of synaptic plasticity.

Detailed description

Grip1 (glutamate receptor interacting protein 1) binds GluR2 (Gria2)-containing α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors and couples them to structural and signaling complexes in neurons. Deletion of both GRIP1 and GRIP2 blocks long-term synaptic depression (LTD) in primary cultures of mouse cerebellar neurons in a model system of synaptic plasticity.

The first PDZ (postsynaptic density protein-95 (PSD-95)/Drosophila disc large tumor suppressor (Dlg1)/zona occludens-1 protein) domain of the Grip1 gene is flanked by loxP sites in this conditional targeted mutant strain. Expression of Cre recombinase results in the deletion of the first PDZ1 exon, causing a frameshift mutation that introduces an immediate stop codon in the GRIP1 transcript.

Development

Two exons encode the first PDZ (postsynaptic density protein-95 (PSD-95)/Drosophila disc large tumor suppressor (Dlg1)/zona occludens-1 protein) domain of the targeted gene. An Frt-flanked PGK neomycin cassette (in reverse transcriptional orientation) followed by a loxP site was introduced to the intron located between the two exons. An additional loxP site was placed 5' of the first PDZ exon. The targeted mutation was created through homologous recombination in (129X1/SvJ x 129S1/Sv)F1- Kitl⁺-derived R1 embryonic stem (ES) cells. Resultant chimeric animals were bred to C57BL/6 mice. Heterozygotes were then bred to C57BL/6 background animals expressing FLP recombinase from a human ACTB promoter (Tg(ACTFLPe)#Dym) to delete the neomycin cassette. This strain was backcrossed to C57BL/6 for 10 generations by the donating laboratory.

Allele

Symbol: Grip1^tm1Rlh
Name: targeted mutation 1, Richard L Huganir
MGI accession ID: MGI:3798437
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Marker symbol: Grip1
Marker name: glutamate receptor interacting protein 1
Chromosome: 10
Strain of origin: (129X1/SvJ x 129S1/Sv)F1-Kitl⁺
Molecular note: A loxP site was inserted upstream of the first coding exon and an frt-flanked neo cassette with a 3' loxP site was inserted downstream of this exon. Germ line, flp-mediated recombination was used to remove the neo cassette leaving the first coding exon floxed.