C57BL/6N-Slc6a4^{em1(flpo)Fuyu}/J

Stock number: 033674
Product name: SERT-ires-Flpo
Research areas: Neurobiology Research, Research Tools

Description

SERT-ires-Flpo CRISPR-derived knock-in mice are designed to have the endogenous Slc6a4 (SERT) promoter/enhancer regions directing expression of optimized FLP recombinase (FLPo) to serotoninergic neurons, without disrupting endogenous Slc6a4 expression. These mice are useful for studying circuit function involving serotonergic neurons, and may also be combined with Cre-lox lines for intersectional studies of sub-populations of serotonergic neurons.

Detailed description

SERT-ires-Flpo CRISPR-derived knock-in mice have an IRES-FLPo cassette inserted downstream of the stop codon of the solute carrier family 6 (neurotransmitter transporter, serotonin), member 4 (Slc6a4) gene. Slc6a4 encodes the sertonin transporter (SERT) which pumps the neurotransmitter serotonin from synaptic spaces into presynaptic neurons. The encoded protein terminates the action of serotonin and recycles it in a sodium-dependent manner. SERT has been implicated in a number of disorders, including alcoholism, clinical depression, obsessive-compulsive disorder (OCD), hypertension and generalized social phobia. The presence of IRES-FLPo cassette downstream of the stop codon allows for expression of SERT and FLPo. Homozygous mice are viable and fertile. When these mice are bred with mice containing frt-flanked sequence, FLPo-mediated recombination will result in deletion of the frt-flanked sequences in serotonergic neurons of the offspring.

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

Development

Plasmids encoding a signal guide RNA designed to introduce an internal ribosomal entry site (IRES) and a Flpo recombinase sequence downstream of the stop codon of the solute carrier family 6 (neurotransmitter transporter, serotonin), member 4 (Slc6a4) gene and the cas9 nuclease were introduced into the cytoplasm C57BL/6NCrl-derived fertilized eggs with well recognized pronuclei. Correctly targeted pups were identified by Southern blot and were further 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: Slc6a4^{em1(flpo)Fuyu}
Name: endonuclease-mediated mutation 1, Fu Yu
MGI accession ID: MGI:6316965
Generation method: Endonuclease-mediated
Attributes: Recombinase-expressing
Marker symbol: Slc6a4
Marker name: solute carrier family 6 (neurotransmitter transporter, serotonin), member 4
Marker synonyms: 5HTT; 5-HTT; 5-HTTLPR; hSERT; Htt; OCD1; SERT; SERT1
Chromosome: 11
Strain of origin: C57BL/6NCrl
Site of expression: Serotonergic neurons
Molecular note: CRISPR/Cas9-system was used to introduce an internal ribosomal entry site (IRES) and a Flpo recombinase sequence downstream of the stop codon of the solute carrier family 6 (neurotransmitter transporter, serotonin), member 4 (Slc6a4) gene.