B6N.129S1-Mrgprb4^{tm4.1(flpo)And}/J

Stock number: 021078
Product name: MrgprB4-EGFPf-2A-FLP-ACN knock-in/knock-out
Research areas: Neurobiology Research, Research Tools

Description

The entire open reading frame of the Mrgprb4 (MAS-related GPR, member B4) gene was replaced with an EGFPf-2A-FLP-ACN cassette in these knockout/knockin allele mutant mice. Constitutive EGFP expression marks Mrgprb4-expressing sensory neurons in the dorsal root ganglion (DRG). When transduced with FLP-dependent adeno-associated virus (AAV), Mrgprb4-specific expression of codon optimized FLP recombinase can be induced.

Detailed description

A rare population of unmyelinated sensory neurons express G protein-coupled receptor Mrgprb4 (MAS-related GPR, member B4). These neurons exclusively innervate hairy skin with large terminal arborizations that resemble the receptive fields of C-tactile afferents in humans.

The entire open reading frame of the gene was replaced by an EGFPf-2A-FLP-ACN cassette in these knockout/knockin mutant allele mice. Constitutive nuclear-localized EGFP expression marks Mrgprb4-expressing sensory neurons in the dorsal root ganglion (DRG). Although not tested, it is anticipated that when transduced with FLP-dependent adeno-associated virus (AAV), Mrgprb4-specific expression of codon optimized FLP recombinase can be induced.

Development

The entire open reading frame, encoded by a single exon, was replaced with an EGFPf-2A-FLP-ACN cassette in 129S1/Sv-p⁺Tyr⁺Kitl ⁺-derived CJ7 embryonic stem (ES) cells.

Farnesylated EGFP (EGFPf) was fused to FLPo (codon optimized FLP recombinase) via foot-and-mouth disease virus 2A peptide (FMDV 2A or F2A) sequence and was ligated to the self-excising loxP-flanked pol-II promoter-neomycin resistance cassette (ACN).

This strain was backcrossed to C57BL/6N for five or more generations by the donating laboratory.

Allele

Symbol: Mrgprb4^{tm4.1(flpo)And}
Name: targeted mutation 4.1, David J Anderson
MGI accession ID: MGI:5448564
Generation method: Targeted
Attributes: Recombinase-expressing
Marker symbol: Mrgprb4
Marker name: MAS-related GPR, member B4
Chromosome: 7
Strain of origin: 129S1/Sv-Oca2⁺ Tyr⁺ Kitl⁺
Expressed genes: GFP,Green Fluorescent Protein,; FLP,FLP recombinase,Saccharomyces cerevisiae
Site of expression: Constitutive nuclear-localized EGFP expression marks Mrgprb4-expressing sensory neurons in the dorsal root ganglion.
Molecular note: The entire open reading frame, encoded by a single exon, was replaced with an EGFPf-2A-FLP-ACN cassette. Farnesylated EGFP (EGFPf) was fused to FLPo (codon optimized FLP recombinase) via foot-and-mouth disease virus 2A peptide (FMDV 2A or F2A) sequence and was ligated to the self-excising loxP-flanked pol-II promoter-neomycin resistance cassette (ACN). Cre-mediated recombination removed the neo cassette.