Overview

This mutant strain carries a floxed allele of the Nr6a1 (nuclear receptor subfamily 6, group A, member 1; also called Gcnf) gene. Cre-mediated recombination can be carried out to create a knockout allele in which exon 7, encoding the beginning of the ligand binding domain, is deleted.

Donating Investigator

David Page, Whitehead Institute for Biomedical Research

Genetic overview

Genetic Background	Generation

Nr6a1^{tm1c(EUCOMM)Wtsi}

Allele Type	Gene Symbol	Gene Name
Targeted (Conditional ready (e.g. floxed))	Nr6a1	nuclear receptor subfamily 6, group A, member 1

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$2,854.50 Domestic price Cryo Recovery

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Details

Detailed Description

Nr6a1 (nuclear receptor subfamily 6, group A, member 1; also called Gcnf) is expressed in fetal ovarian cells, peaking at embryonic day 14.5 (E14.5). Both somatic stem cells and embryonic stem cells require NR6A1 to silence the activities of Pou5f1 (POU domain, class 5, transcription factor 1; also called Oct4) and Nanog (Nanog homeobox) in the transition from pluripotent to differentiated cell state. Interestingly, the gene is not required for this process in fetal ovarian germ cells and it is not required for initiation of meiosis and oogenesis.

Exon 7 of the Nr6a1 gene, encoding the beginning of the ligand binding domain, is flanked by loxP sites in this mutant strain. Homozygous floxed animals are phenotypically normal for gross morphology, behavior, fertility, and viability.

Cre-mediated recombination can be carried out to produce a knockout allele in which exon 7 is deleted. Crosses with tamoxifen-inducible UBC-cre/ERT2 mice (e.g. Stock No. 008085) produce a widespread knockout of the gene. Homozygous knockout embryos die prenatally and display reduced size, impaired neural tube closure, and failure to turn from the lordotic to the fetal position.

Development

This floxed allele of Nr6a1 (called Gcnf) is derived from embryonic stem (ES) cells made by the EUCOMM (European Conditional Mouse Mutagenesis) Project, Wellcome Trust Sanger Institute. The original targeting vector incorporated a cassette containing a splice acceptor (SA) and internal ribosome entry site (IRES) upstream of a lacZ reporter gene followed by a polyadenylation (pA) signal. A selection cassette followed, consisting of a neomycin resistance gene (neo) driven by an autonomous promoter (hBactP) and pA signal. Both cassettes, separated by a loxP site, were flanked by FRT sites within the intron 6 region. Additionally, loxP sites were introduced on either side of exon 7 which encodes the ligand-binding domain. The mutation was created in C57BL/6NTac-derived JM8.F6 embryonic stem (ES) cells. The original targeted allele is reflected in Stock No. 023804.

After Flp-mediated recombination through crosses with ACTB-FLPe mice (e.g. Stock No. 005703), the IRES-lacZ and neomycin cassettes were removed, leaving exon 7 of the gene flanked by loxP sites. This strain was maintained on a C57BL/6NTac genetic background by the donating laboratory. Upon arrival at The Jackson Laboratory, mice were rederived with C57BL/6NJ (see Stock No. 005304)

In 2014, a 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the first generation rederived living colony at The Jackson Laboratory Repository. While the 27 markers throughout the genome suggested a C57BL/6 genetic background, 3 of 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating in some mice (chromosomes 8 [~15.2 Mbp], 13 [~41.0 Mbp] and 15 [~57 Mbp]). These data suggest that the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6J;C57BL/6N genetic background.

Control Suggestions

Approximate Controls

Additional Information

Considerations for Choosing Controls

Selected References

Skarnes WC; Rosen B; West AP; Koutsourakis M; Bushell W; Iyer V; Mujica AO; Thomas M; Harrow J; Cox T; Jackson D; Severin J; Biggs P; Fu J; Nefedov M; de Jong PJ; Stewart AF; Bradley A. 2011. A conditional knockout resource for the genome-wide study of mouse gene function. Nature 474(7351):337-42PubMed: 21677750 MGI: J:173534

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Genetics

Nr6a1^{tm1c(EUCOMM)Wtsi}

Allele Symbol: Nr6a1^{tm1c(EUCOMM)Wtsi}

Allele Name	targeted mutation 1c, Wellcome Trust Sanger Institute
Allele Type	Targeted (Conditional ready (e.g. floxed))
Allele Synonym(s)	Nr6a1^{tm1c(EUCOMM)Wtsi}; targeted mutation 1c, Wellcome Trust Sanger Institute
Gene Symbol and Name	Nr6a1, nuclear receptor subfamily 6, group A, member 1
Gene Synonym(s)	NR61; GCNF; GCNF1; CT150; hGCNF; 1700113M01Rik; Gcnf; hRTR; NCNF; RTR; 1700113M01Rik; germ cell nuclear factor; RIKEN cDNA 1700113M01 gene; Gcnf
Strain of Origin	C57BL/6N
Chromosome	2
General Note	Cell line EPD0105_6_D03 was successfully used to make chimeric mice. Germline transmission was accomplished. J:101977
Molecular Note	The L1L2_Bact_P cassette was inserted at position 38743429 of Chromosome 2 upstream of the critical exon 7 (Build GRCm38). The cassette is composed of an FRT site followed by lacZ sequence and a loxP site. This first loxP site is followed by neomycin resistance gene under the control of the human beta-actin promoter, SV40 polyA, a second FRT site and a second loxP site. A third loxP site is inserted downstream of the targeted exon 7 at position 38742577. The critical exon 7 is thus flanked by loxP sites. A "conditional ready" (floxed) allele can be created by flp recombinase expression in mice carrying this allele. Subsequent cre expression results in a knockout mouse. If cre expression occurs without flp expression, a reporter knockout mouse will be created. Further information on targeting strategies used for this and other IKMC alleles can be found at http://www.informatics.jax.org/mgihome/nomen/IKMC_schematics.shtml. Flp-mediated recombination removed the reporter and neomycin resistance cassette and left exon 7 floxed.

Disease/Phenotype

Disease Terms

Research Areas By Genotype

This mouse can be used to support research in many areas including:

Research Tools
- Cre-lox System
  - loxP-flanked Sequences
- Developmental Biology Research
  - Cre-lox System

Mammalian Phenotype Terms by Genotype

The following phenotype information is associated with a similar, but not exact match to this JAX^® Mice strain

Genotype: Nr6a1^{tm1c(EUCOMM)Wtsi}/Nr6a1^{tm1c(EUCOMM)Wtsi} Ndor1^{Tg(UBC-cre/ERT2)1Ejb}/0
involves: 129S/SvEv * C57BL/6 * C57BL/6N * SJL

mammalian phenotype

reproductive system phenotype
- Normal - tamoxifen-treated mice exhibit normal early meiosis and oogenesis at E16.5

References

Skarnes WC; Rosen B; West AP; Koutsourakis M; Bushell W; Iyer V; Mujica AO; Thomas M; Harrow J; Cox T; Jackson D; Severin J; Biggs P; Fu J; Nefedov M; de Jong PJ; Stewart AF; Bradley A. 2011. A conditional knockout resource for the genome-wide study of mouse gene function. Nature 474(7351):337-42PubMed: 21677750 MGI: J:173534

Additional - Nr6a1^{tm1c(EUCOMM)Wtsi} related

Technical Support

Contact Technical Support

Genotyping Protocols
- Standard PCR: Nr6a1^{tm1c(EUCOMM)Wtsi}
- Genotyping resources and troubleshooting
Breeding Considerations

Heterozygous and homozygous floxed mice are viable and fertile.
- Additional Breeding and Husbandry Support
Citation
When using the B6(Cg)-Nr6a1^{tm1c(EUCOMM)Wtsi}/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #023803 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Pricing & Availability

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Genotype

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Cryorecovery - Not-For-Profit & Academic Pricing

Service	Genotype	Price
	Heterozygous for Nr6a1<tm1c(EUCOMM)Wtsi>

We will fulfill your order by providing at least two carriers for each strain ordered. The total number, sex, and genotypes provided will vary, although typically 8 or more animals are provided. Please check genotypes which will be recovered. While the genotypes of all animals produced will be communicated to you prior to scheduling shipment, the genotypes of animals provided may not reflect the mating scheme and genotypes described in the strain description. Animals are typically ready to ship in 11-14 weeks. If a second recovery is required to produce the minimum number of animals, then delivery time would increase to approximately 25 weeks. If we fail to produce animals of the correct genotype, you will not be charged. We cannot guarantee the reproductive success of mice shipped to your facility. If the mice are lost after the first three days (post-arrival) or do not produce progeny at your facility, a new order and fee will be necessary.

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The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project.

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Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Control Suggestions

Approximate Controls

Additional Information

Selected References

Nr6a1tm1c(EUCOMM)Wtsi

Allele Symbol: Nr6a1tm1c(EUCOMM)Wtsi

Disease Terms

Research Areas By Genotype

This mouse can be used to support research in many areas including:

Mammalian Phenotype Terms by Genotype

Genotype: Nr6a1tm1c(EUCOMM)Wtsi/Nr6a1tm1c(EUCOMM)Wtsi Ndor1Tg(UBC-cre/ERT2)1Ejb/0involves: 129S/SvEv * C57BL/6 * C57BL/6N * SJL

mammalian phenotype

References

Additional - Nr6a1tm1c(EUCOMM)Wtsi related

Genotyping Protocols

Breeding Considerations

Citation

Animal Health Reports

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Live Mouse

Cryorecovery - Not-For-Profit & Academic Pricing

Related Products and Services

Payment Terms and Conditions

The Jackson Laboratory's Genotype Promise

Terms of Use

Additional Use Restrictions Apply

Licensing Information

JAX® Mice, Products & Services Conditions of Use

No Warranty

Credit for PRODUCTS or SERVICES

Animal Care and Use for SERVICES

No Liability

Nr6a1^{tm1c(EUCOMM)Wtsi}

Allele Symbol: Nr6a1^{tm1c(EUCOMM)Wtsi}

Genotype: Nr6a1^{tm1c(EUCOMM)Wtsi}/Nr6a1^{tm1c(EUCOMM)Wtsi} Ndor1^{Tg(UBC-cre/ERT2)1Ejb}/0
involves: 129S/SvEv * C57BL/6 * C57BL/6N * SJL

Additional - Nr6a1^{tm1c(EUCOMM)Wtsi} related