B6N(Cg)-Slc6a3^{tm1.1(flpo)Fuyu}/J

Stock number: 033673
Product name: DAT-ires-Flpo
Research areas: Neurobiology Research, Research Tools

Description

DAT-ires-Flpo knock-in mice are designed to have the endogenous Slc6a3 (DAT) promoter/enhancer regions directing expression of optimized FLP recombinase (FLPo) to dopaminergic neurons, without disrupting endogenous Slc6a3 expression. These mice are useful for studying circuit function involving dopaminergic neurons, and may also be combined with Cre-lox lines for intersectional studies of sub-populations of dopaminergic neurons.

Detailed description

Slc6a3 encodes the dopamine transporter (DAT) which pumps the neurotransmitter dopamine out of the synapse back into cytosol. Dopamine release in the ventral tegmental area (VTA) plays a major role in reward-motivated behavior. Defects in dopamine transport reduces the rate of transport 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. Cocaine is able to block this transport, by binding directly to the DAT, increasing reward-related dopamine activity.

DAT-ires-Flpo knock-in mice have an IRES-FLPo cassette inserted downstream of the stop codon of the solute carrier family 6 (neurotransmitter transporter, dopamine), member 3 (Slc6a3) gene. Homozygous mice are viable and fertile. The presence of IRES-FLPo cassette downstream of the stop codon allows for expression of DAT and FLPo. When these mice are bred with mice containing frt-flanked sequence, FLPo-mediated recombination will result in deletion of the frt-flanked sequences in dopaminergic neurons of the offspring.

These mice are useful for studying circuit function involving dopaminergic neurons, and may also be combined with Cre-lox lines for intersectional studies of sub-populations of dopaminergic neurons.

Development

The DAT-ires-Flpo targeting construct was designed to insert a loxN-flanked neomycin resistance (neo) cassette followed by an internal ribosomal entry site (IRES) and a Flpo recombinase sequence downstream of the stop codon of the solute carrier family 6 (neurotransmitter transporter, dopamine), member 3 (Slc6a3) gene. This construct was electroporated into C57BL/6NCrl embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and the resulting chimeric males were bred to CMV-cre mice to remove the neo cassette. Offspring were crossed with C57BL/6NCrl mice to remove the cre-expressing transgene, and resulting DAT-ires-Flpo mice were bred to C57BL/6NCrl mice for at least five generations. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6NJ oocytes (Stock No. 005304).

Allele

Symbol: Slc6a3^{tm1.1(flpo)Fuyu}
Name: targeted mutation 1.1, Fu Yu
MGI accession ID: MGI:6316964
Generation method: Targeted
Attributes: Recombinase-expressing
Marker symbol: Slc6a3
Marker name: solute carrier family 6 (neurotransmitter transporter, dopamine), member 3
Marker synonyms: DAT; Dat1; PKDYS; PKDYS1
Chromosome: 13
Strain of origin: C57BL/6NCrl
Site of expression: Dopaminergic neurons
Molecular note: The DAT-ires-Flpo targeting construct was designed to insert a loxN-flanked neomycin resistance (neo) cassette followed by an internal ribosomal entry site (IRES) and a Flpo recombinase sequence downstream of the stop codon of the solute carrier family 6 (neurotransmitter transporter, dopamine), member 3 (Slc6a3) gene. Correctly targeted ES cells were injected into blastocysts and the resulting chimeric males were bred to CMV-cre mice to remove the neo cassette.