B6;129S4-Plxnd1^{tm2.1(flpo)Zjh}/J

Stock number: 036295
Product name: PlexinD1-2A-flpO
Research areas: Research Tools

Description

PlexinD1-2A-flpO knock-in mice express flpO (mouse codon-optimized Flp recombinase) from the mouse Plxnd1 (plexin D1) promoter in PLXND1-positive cortical glutamatergic pyramidal neurons (PyNs).

Detailed description

PlexinD1-2A-flpO knock-in mice express flpO (mouse codon-optimized Flp recombinase) from the Plxnd1 (plexin D1) promoter/enhancer elements. As such, when PlexinD1-2A-flpO mice are bred with mice containing FRT-flanked sequence(s), flpO-mediated recombination will result in deletion of the FRT'ed sequence(s) in PLXND1-positive cortical glutamatergic pyramidal neurons (PyNs).

Endogenous Plxnd1 gene expression is not altered by the knock-in insert. Homozygous mice are viable and fertile.

A set of temporally inducible mouse Cre and Flp knock-in driver lines that enable combinatorial targeting of major progenitor types and projection classes of glutamatergic pyramidal neurons (PyNs) and their progenitors were described in Matho et al, 2020 (BioRxiv). These gene knock-in's recapitulate the endogenous spatial, temporal and cellular expression patterns of transcription factors and effector genes. Several of these strains are available through The Jackson Laboratory:

Driver Lines:
Lhx2-2A-CreER 036293
PlexinD1-2A-CreER 036294
PlexinD1-2A-flpO 036295
Fezf2-2A-CreER 036296
Fezf2-2A-flpO 036297
Tle4-2A-CreER 036298
Tbr1-2A-CreER 036299
Cux1-2A-CreER 036300
Tbr2-2A-CreER 036301
Tis21-2A-CreER 036303
Adcyap1-2A-CreER 033999
Tcerg1l-2A-CreER 034000

Reporter Line:
4 Dual-tTA 036304

Development

A targeting vector was designed to insert flpO (mouse codon-optimized Flp recombinase) and a T2A self-cleaving peptide sequence followed by a loxP-flanked neo cassette upstream of the stop codon of the Plxnd1 (plexin D1) gene.

This construct was electroporated into (C57BL/6 x 129S4/SvJae)F1-derived v6.5 embryonic stem (ES) cells. The neomycin cassette was excised through crosses with CMV-Cre (Stock No. 006054). The resultant PlexinD1-2A-flpO allele was maintained on a mixed C57BL/6 and 129 genetic background by the donating laboratory. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

Allele

Symbol: Plxnd1^{tm2.1(flpo)Zjh}
Name: targeted mutation 2.1, Z Josh Huang
MGI accession ID: MGI:6720835
Generation method: Targeted
Attributes: Recombinase-expressing
Synonyms: PlexinD1-2A-flpO; PlexinD1-flpO
Marker symbol: Plxnd1
Marker name: plexin D1
Marker synonyms: 6230425C21Rik; b2b1863Clo; b2b553Clo; CHTD9; PLEXD1
Chromosome: 6
Strain of origin: (C57BL/6 x 129S4/SvJae)F1
Molecular note: The allele was generated by inserting a 2A-flpo expression cassette in-frame before the STOP codon of the Plxnd1 gene. The targeting vectors generated using BAC clones from the RPCI-23&24 library were electroporated into a 129SVj/B6 hybrid ES cell line (v6.5). The Resulting ES cell clones were used for tetraploid complementation to obtain male heterozygous mice. The neo selection cassette was excised by crossing to cre expressing mice.