B6.Cg-Slc6a3^tm1.1Asor/J

Stock number: 029381
Product name: DAT^HA
Research areas: Neurobiology Research, Research Tools

Description

DAT^HA mice express an HA tagged dopamine receptor (Slc6a3) gene. They may be useful for studying endogenous dopamine transporter trafficking.

Detailed description

DAT^HA mice express an HA tagged solute carrier family 6 (neurotransmitter transporter, dopamine), member 3 (Slc6a3) gene. Slc6a3 encodes the dopamine transporter (DAT) which pumps the neurotransmitter dopamine out of the synapse back into dopaminergic neurons. Dopamine release in the striatum plays a major role in reward-motivated behavior. Defects in dopamine re-uptake reduce the rate of dopamine clearance from the synapse, thus increasing the levels of dopamine in the brain. DAT has been implicated in a number of dopamine-related disorders, including attention deficit hyperactivity disorder, bipolar disorder, clinical depression, and alcoholism. The HA tag is inserted into the 5’ end of exon 4 which encodes the second extracellular loop of dopamine transporter, allowing for detection in living neurons in culture and brain slices by HA antibodies. Homozygous mice are viable and fertile and express normal levels of DAT.

Development

A targeting construct was designed to insert an HA tag at the 5’ end of exon 4 of the solute carrier family 6 (neurotransmitter transporter, dopamine), member 3 ( Slc6a3) gene. A loxP-flanked neomycin resistance (neo) cassette was inserted in intron 3. This construct was electroporated into (C57BL/6J x 129X1/SvJ)F1-derived EC7.1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts and the resulting chimeric animals were crossed to Actb-cre mice on a C57BL/6 background to remove the neo selection cassette. These DAT^HA mice were bred together to remove the Cre-expressing transgene and were subsequently bred to C57BL/6 mice for at least 9 generations. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J (Stock No. 000664) oocytes.

Allele

Symbol: Slc6a3^tm1.1Asor
Name: targeted mutation 1.1, Alexander Sorkin
MGI accession ID: MGI:5319237
Generation method: Targeted
Attributes: Reporter
Marker symbol: Slc6a3
Marker name: solute carrier family 6 (neurotransmitter transporter, dopamine), member 3
Chromosome: 13
Strain of origin: (C57BL/6J x 129X1/SvJ)F1
Expressed genes: HA,influenza hemagglutinin,; Slc6a3,solute carrier family 6 (neurotransmitter transporter, dopamine), member 3,mouse, laboratory
Molecular note: A floxed neo cassette was inserted downstream of exon 3. Exon 4 was replaced with one with a human influenza hemagglutinin (HA) tag at the 5' end of the exon. Cre-mediated recombination removed the neo cassette. Western blot analysis confirmed normal protein expression levels.