B6.129S7(FVB)-Dbh^{tm1.1(flpo)Pjen}/J

Stock number: 033952
Product name: Dbh^Flpo KI
Research areas: Developmental Biology Research, Research Tools

Description

Dbh^FLPo knock-in/knock-out mice express FLPo recombinase from the endogenous Dbh locus, with FLPo recombinase activity detected in noradrenergic neurons.

Detailed description

These Dbh^Flpo knock-in mice express FLPo recombinase from the endogenous Dbh (dopamine beta hydroxylase) locus, directing FLPo recombinase activity in noradrenergic neurons. The insertion of FLPo abolishes Dbh expression. Homozygotes are not viable (embryonic lethal) due to loss of norepinephrine synthesis.

When bred to the RC::FrePe reporter line, FLPo recombinase activity is detected in norepinephrine and epinephrine nuclei of the brainstem (noradrenergic and adrenergic neurons), and the peripheral nervous system (adrenal medulla, sympathetic ganglia, cranial ganglia, and enteric neurons). FLPo recombinase activity is also detected in E12.5 embryos. Mice that are heterozygous for the targeted mutation are viable and fertile.

Development

A targeting vector containing a Flpo-poly(A) cassette and a loxP-flanked PGK/Em7-Neo cassette was used to replace 107 bp of exon 1 of the Dbh gene. The construct was electroporated into 129S7/SvEvBrd-Hprt^b-m2 derived AB2.2 embryonic stem (ES) cells. Correctly targeted ES cells were injected into B6(Cg)-Tyr^c-2J/J blastocysts. The resulting chimeric animals were tested for germline transmission and bred to C57BL/6J mice. Heterozygous offspring were crossed to FVB/N-Tmem163^{Tg(ACTB-cre)2Mrt}/J mice (Stock No. 003376) to remove the loxP-flanked Neo cassette, and the line was subsequently maintained by backcrossing to C57BL/6J mice (Stock No. 000664) for 27 generations. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes.

Allele

Symbol: Dbh^{tm1.1(flpo)Pjen}
Name: targeted mutation 1.1, Patricia Jensen
MGI accession ID: MGI:6358815
Generation method: Targeted
Attributes: Recombinase-expressing
Synonyms: Dbh^Flpo
Marker symbol: Dbh
Marker name: dopamine beta hydroxylase
Marker synonyms: DBM; DOPBHY; ORTHYP1
Chromosome: 2
Strain of origin: 129S7/SvEvBrd-Hprt1^b-m2
Site of expression: Noradrenergic neurons.
Molecular note: A targeting vector containing a Flpo-poly(A) cassette and a loxP-flanked PGK/Em7-Neo cassette was used to replace 107 bp of exon 1 of the gene. Cre-mediated recombination removed the floxed neo cassette.
General note: The version of this allele that contains the neomycin selection cassette, Dbh^{tm1(flpo)Pjen}, has been analyzed.