B6.129(C)-Scn9a^tm2.1Jwo/J

Stock number: 035964
Product name: Nav1.7^TAP
Research areas: Neurobiology Research, Research Tools

Description

Nav1.7^TAP mice have a TAP-tag inserted into the open reading frame at the 3'-end, prior to the stop codon in exon 27, of the Scn9a gene. Nav1.7 plays an important role in pain, nociception, and opioid signaling, making these mice useful for mapping protein interactions of Nav1.7 during chronic and acute pain responses.

Detailed description

Nav1.7^TAP mice have a TAP-tag inserted into the open reading frame at the 3'-end, prior to the stop codon in exon 27, of the sodium channel, voltage-gated, type IX, alpha (Scn9a) gene. The TAP-tag contains a HAT domain, a TEV cleavage site, and 3x FLAG-tags. Nav1.7 plays an important role in pain, nociception, and opioid signaling. Mutations in SCN9A have been associated with different "channelopathies" with electrical hyperactivity of sensory neurons in dorsal root and a low reactivity of sympathetic ganglia neurons.

Homozygous Nav1.7^TAP mice exhibit no deficit in motor function, and they have normal acute pain behavior. Using immunohistochemical mean to look at expression, the TAP-tag is expressed in neurons of the olfactory bulb and the medial amygdala, along with the hypothalamus, midbrain, hindbrain, and the spinal cord. By Western blot, FLAG-tag expression is present in the olfactory bulb, hypothalamus, spinal cord, sciatic nerve and DRG, but not detectable in cortex, cerebellum, skin, lung, heart and pancreas. Homozygous mice are viable and fertile.

Development

The Nav1.7^TAP targeting construct was designed to insert a TAP-tag into the open reading frame at the 3'-end, prior to the stop codon in exon 27, of the sodium channel, voltage-gated, type IX, alpha (Scn9a) gene. The TAP-tag contains a HAT domain, a TEV cleavage site, and 3x FLAG-tags, and is followed by a loxP-flanked neo cassette. This construct was electroporated into 129/Sv-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts and the chimeric offspring were bred to CMV-Cre mice (Stock No. 006054) to remove the neo cassette. The resulting mice were crossed to remove the cre-expressing transgene and subsequently Nav1.7^TAP mice were bred to C57BL/6J mice for at least 10 generations. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

Allele

Symbol: Scn9a^tm2.1Jwo
Name: targeted mutation 2.1, John N Wood
MGI accession ID: MGI:6147829
Generation method: Targeted
Attributes: No functional change; Epitope tag
Synonyms: NaV1.7^TAP
Marker symbol: Scn9a
Marker name: sodium channel, voltage-gated, type IX, alpha
Marker synonyms: ETHA; FEB3B; GEFSP7; HSAN2D; Nav1.7; NENA; NE-NA; PN1; Scn2a; SFNP
Chromosome: 2
Strain of origin: 129
Molecular note: A TAP-tag containing a HAT domain, a TEV cleavage site and 3x FLAG-tags and a floxed neo cassette were inserted into the open reading frame at the 3'-end prior to the stop codon in exon 27. Cre mediated recombination removed the neo cassette.