B6N.129S1-Mrgprb4^tm3(cre)And/J

Stock number: 021077
Research areas: Neurobiology Research, Research Tools, Sensorineural Research

Description

The entire open reading frame of the Mrgprb4 (MAS-related GPR, member B4) gene was replaced by an mtdTomato-2A-NLSCre-Frt-PGK-Neomycin-FRT cassette in these knockout/knockin allele mutant mice. When virally transduced, they are useful in studies of Mrgprb4-expressing sensory neurons. Constitutive expression of mtdTomato has been demonstrated in dorsal root ganglion (DRG) cells.

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 mtdTomato-2A-NLSCre-Frt-PGK-Neomycin-FRT cassette in these knockout/knockin allele mutant mice. Mutant mice have no phenotype on their own, but when virally transduced (e.g. using Cre-dependent adeno-associated virus (AAV) to express genetically encoded calcium sensor GCaMP3.0 or hm3(G_q-coupled)DREADD) they are useful in studies of Mrgprb4-expressing sensory neurons. Constitutive Mrgprb4-specific expression of mtdTomato has been demonstrated in dorsal root ganglion (DRG) cells.

Development

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

The membrane-tagged tdTomato (mtdTomato) contains the 8 amino acids of the MARCKS sequence (MGCCFSKT) fused to the N-terminus of the full-length tdTomato and was connected to a nuclear localization signal (NLS)-tagged Cre recombinase via an intervening foot-and-mouth disease virus 2A peptide (FMDV 2A or F2A) sequence.

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

Allele

Symbol: Mrgprb4^tm3(cre)And
Name: targeted mutation 3, David J Anderson
MGI accession ID: MGI:5448562
Generation method: Targeted
Attributes: Recombinase-expressing
Synonyms: MrgprB4-mtdTomato-2A-NLScre-frt-PGK-neo-frt; Mrgprb4-cre
Marker symbol: Mrgprb4
Marker name: MAS-related GPR, member B4
Chromosome: 7
Strain of origin: 129S1/Sv-Oca2⁺ Tyr⁺ Kitl⁺
Expressed genes: cre,cre recombinase,bacteriophage P1
Site of expression: Constitutive Mrgprb4-specific expression of mtdTomato has been demonstrated in dorsal root ganglion cells.
Molecular note: The entire open reading frame, encoded by a single exon, was replaced with an mtdTomato-2A-NLSCre-Frt-PGK-Neomycin-FRT cassette. The membrane-tagged tdTomato (mtdTomato) contains the 8 amino acids of the MARCKS sequence (MGCCFSKT) fused to the N-terminus of the full-length tdTomato and was connected to a nuclear localization signal (NLS)-tagged Cre recombinase via an intervening foot-and-mouth disease virus 2A peptide (FMDV 2A or F2A) sequence.