STOCK Ndufs2^tm1.1Job Tg(CAG-cre/Esr1*)5Amc/J

Stock number: 038571
Product name: Inducible MCI-PARK
Strain synonyms: Ndufs2^fl;CAGGCre-ER^TM , ESR-NDUFS2
Research areas: Neurobiology Research, Research Tools

Description

Removal of this mouse colony is imminent. If live mice are needed for your studies, it is advised that they be ordered immediately. After removal, the mice will be available from a cryorecovery.

Inducible MCI-PARK mice, based on the ESR-NDUFS2 strain described by Fernández-Agüera, 2015 (PMID: 26437605), were generated through an intercross of mice carrying a floxed allele of mouse NADH:ubiquinone oxidoreductase core subunit S2 (Ndufs2; derived from Stock No. 036313) with animals that broadly express tamoxifen-inducible Cre recombinase (Tg(CAG-cre/Esr1*)5Amc; Stock No. 004682). Induction with tamoxifen creates a conditional knock-out of Ndufs2, useful in studies of Parkinson's disease (PD).

Distribution of this double mutant strain (Stock No. 038571) was supported by the Michael J. Fox Foundation for Parkinson's Research.

Detailed description

Inducible MCI-PARK mice, based on the ESR-NDUFS2 strain described in Fernández-Agüera, 2015 (PMID: 26437605), were generated through an intercross of mice carrying an allele of the mouse Ndufs2 gene in which exon 2 is flanked by loxP sites (derived from Stock No. 036313) with animals that broadly express tamoxifen-inducible Cre recombinase (Tg(CAG-cre/Esr1*)5Amc; Stock No. 004682). Induction with tamoxifen creates a conditional knock-out of the mouse Ndufs2 gene useful in studies of Parkinson's disease (PD).

Mice that are heterozygous or homozygous for the Ndufs2 floxed allele are viable and fertile. Mice hemizygous for the Tg(CAG-cre/Esr1*)5Amc transgene are viable and fertile, but homozygotes are non-viable.

Distribution of this double mutant strain (Stock No. 038571) was supported by the Michael J. Fox Foundation for Parkinson's Research.

Development

Mice carrying the Ndufs2^tm1.1Job allele (derived from Stock No. 036313) were intercrossed with mice expressing Tg(CAG-cre/Esr1*)5Amc (Stock No. 004682) at The Jackson Laboratory (2023). Resultant mice are on a mixed genetic background that likely involves C57BL/6J, C57BL/6N, CBA/J, SWR, 129X1/SvJ, 129S1/Sv, and SJL.

Sperm from compound mutant mice, commonly referred to as Inducible MCI-PARK, was cryopreserved. Cryorecovery/live colony development will involve C57BL/6J (Stock No. 000664).

The Ndufs2^tm1.1Job (Ndufs2^fl) allele (see Stock No. 036313), made by Dr. José López-Barneo (Universidad de Sevilla), was created by homologous recombination in (129X1/SvJ x 129S1/Sv)F1- Kitl⁺-derived R1 embryonic stem (ES) cells. A loxP site was placed in intron 1 and a loxP-FRT-neomycin-FRT cassette was introduced to intron 2 of the mouse Ndufs2 gene. The neomycin cassette was excised from the resultant mice through crosses with B6;SJL-Tg(ACTFLP3)9205Dym/J mice (Stock No. 003800), leaving exon 2 flanked by loxP sites.

The Tg(CAG-cre/Esr1*)5Amc (pCAGGCre-ER^TM) transgene (see Stock No. 004682) was designed by Drs. Shigemi Hayashi and Andrew P. McMahon (while at Harvard University) to have the CMV-IE enhancer/chicken β-actin/rabbit β-globin hybrid promoter (CAG; originally from the pCAGGS vector) and CreER^TM fusion gene (CreER^TM; Cre recombinase fused to a G525R mutant form of the mouse estrogen receptor ligand binding domain). The transgene was introduced into B6CBF1 donor eggs. The resulting male transgenic mice were first backcrossed for two generations on the SWR background. Next, mice were backcrossed onto the C57BL/6J background for five generations.

Allele

Symbol: Tg(CAG-cre/Esr1*)5Amc
Name: transgene insertion 5, Andrew P McMahon
MGI accession ID: MGI:2182767
Generation method: Transgenic
Attributes: Recombinase-expressing; Inducible
Synonyms: CAG-CreEr; CAGG-Cre; CaggCreER; CAGGCre-ER; CAGGCre-ER^TM; CAGGCre-ERtm line 5.8; CAGGS-CreER; CAGGS-CreErT; CMV-creERT; Cre-ER; Cre-ER^TM; CreESR^T; ER-cre; Tg(CAG-cre/Esr1)5Amc; Tg(cre/Esr1)5Amc; Tg:Cag-Cre/Esr1; Tg^CreER; TgCAGGCreER; uCreER^T; CAG-Cre
Marker symbol: Tg(CAG-cre/Esr1*)5Amc
Marker name: transgene insertion 5, Andrew P McMahon
Marker synonyms: CAG-CreERT2; CAGGCre-ER; CAGGCre-ER^TM; CAGG-CreERT2; CAGGCre-ERtm line 5.8; CAGGS-CreER; CAGGS-CreErT; CMV-creERT; Cre-ER; Cre-ERTM; ER-cre; Tg(CAG-cre/Esr1)5Amc; Tg(cre/Esr1)5Amc
Chromosome: UN
Strain of origin: (C57BL/6 x CBA)F1
Expressed genes: cre/Esr1,Cre recombinase and estrogen receptor 1 fusion gene,
Site of expression: tamoxifen-inducible cre; widespread pattern of expression; tamoxifen administration will also induce Cre recombination in developing embryos of treated mothers and in cultured cells derived from transgenic mice
Molecular note: This transgene expresses a fusion protein consisting of Cre recombinase joined to the ligand-binding domain of a mouse estrogen receptor modified to bind to 4-hydroxytamoxifen, but not to endogenous estrogen. The CAG promoter, containing a chicken beta actin promoter/enhancer coupled with the cytomegalovirus immediate-early (CMV-IE) enhancer, drives high levels of expression in most tissues. In the presence of tamoxifen, the fusion protein is transported into the nucleus, where cre can excise loxP-flanked segments from conditionally modified genes.
General note: Homozygous transgenic mice are not viable or fertile. Hemizygous transgenic mice are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities.

In transgenic mice the mutant mouse estrogen receptor does not bind natural ligand at physiological concentrations but will bind the synthetic ligand, 4-hydroxytamoxifen. Restricted to the cytoplasm, the Cre/Esr1 fusion protein can only gain access to the nuclear compartment after exposure to tamoxifen. When crossed with a strain containing loxP sites flanking a sequence of interest, tamoxifen induced, Cre-mediated targeted deletions are generated in the offspring. Tamoxifen administration will induce Cre recombination in developing embryos of treated mothers and in cultured cells derived from transgenic mice.

Note that this transgene does not contain the cre/ERT2 fusion but was mistakenly referred to by the synonym CAG-CreERT2 in publication. J:136965

Note that this transgene does not contain the cre/ERT2 fusion but was mistakenly referred to by the synonym CAGG-CreERT2 in publication. J:130851

Allele

Symbol: Ndufs2^tm1.1Job
Name: targeted mutation 1, Jose Lopez-Barneo
MGI accession ID: MGI:6358287
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed)
Synonyms: Ndufs2^F
Marker symbol: Ndufs2
Marker name: NADH:ubiquinone oxidoreductase core subunit S2
Marker synonyms: CI-49; LHONAR2; MC1DN6; MGC:27667
Chromosome: 1
Strain of origin: Not Specified
Molecular note: A loxP site was inserted upstream of exon 2 and a FRT flanked neomycin cassette with a 3' loxP site was inserted downstream of exon 2. The neo cassette was removed to produce the final conditional allele.