Strain Name:
B6J.129S6-Nkx2-1tm1Ikono/Mmjax
Stock Number:
066764-JAX
Citation ID:
RRID:MMRRC_066764-JAX
Other Names:
Nkx2-1-GFP

Strain Information

Nkx2-1tm1Ikono
Name: NK2 homeobox 1; targeted mutation 1, Laertis Ikonomou
Synonyms: Nkx2-1GFP
Type: Allele
Species: Mus musculus (mouse)
Chromosome: 12
Alteration at locus: Knock-In
Nkx2-1
Name: NK2 homeobox 1
Synonyms: thyroid-specific enhancer-binding protein, T/EBP, tinman, Ttf-1, thyroid transcription factor-1, Titf1
Type: Gene
Species: Mouse
Chromosome: 12
Alteration at locus: Knock-In
NCBI: 21869
Homologene: 2488
Genetic Alterations
Targeting of the eGFP gene to one allele of the endogenous Nkx2-1 locus, replacing endogenous sequences from the second ATG start site through the end of the Nkx2-1 homeobox (positions 7957–9480 in GenBank locus MMU19755).
Genotype Determination
ES Cell Line
W4 derived from 129S6/SvEvTac
Phenotype
Heterozygous: Subclinical Hypothyroidism
Homozygous: Lethal at P0
MeSH Terms
  • Animals
  • Bone Morphogenetic Proteins/antagonists & inhibitors
  • Bone Morphogenetic Proteins/metabolism
  • Cell Separation
  • Embryonic Stem Cells/cytology
  • Embryonic Stem Cells/metabolism
  • Fibroblast Growth Factors/antagonists & inhibitors
  • Fibroblast Growth Factors/metabolism
  • Lung/cytology
  • Lung/embryology
  • Lung/metabolism
  • Mice
  • Mice, Transgenic
  • Signal Transduction/physiology
  • Thyroid Gland/cytology
  • Thyroid Gland/embryology
  • Thyroid Gland/metabolism
  • Tissue Scaffolds
Strain Development
A cDNA insert encoding the GFP reporter gene was delivered by homologous recombination in W4/129S6 ES cells (Taconic Farms, Inc.), replacing endogenous Nkx2-1-encoding sequences from the second ATG start site through the end of the homeobox (positions 7957-9480 in GenBank locus MMU19755). To enable gene targeting, a targeting vector backbone, TV-CD8-Puro (generous gift of Drs. Gordon Keller and Paul Gadue, Mt Sinai School of Medicine, New York), was adapted by conventional cloning as follows: gDNA extracts from 129/Ola ES cells were used as templates for PCR cloning of a short arm of homology, representing 1.9 kb of sequence 5’ to the Nkx2-1 ATG start site (position 7957), and a long arm of homology, representing 10.1 kb of sequence 3’ of Nkx2-1 position 9480. Each arm of homology was cloned into the TV-CD8-Puro backbone and the CD8 cassette was replaced with a cassette encoding enhanced green fluorescence protein (hereafter GFP). The resulting targeting vector was delivered to W4/129S6 ES cells by electroporation. 300 puromycin resistant ES cell colonies were picked and screened by PCR to identify 3 candidate clones. To confirm successful targeting of the Nkx2-1 locus, BclI-digested gDNA extracts were analyzed by Southern blotting after hybridization with a 32P-labeled Nkx2-1 probe, confirming the introduction of the new BclI restriction site in the expected location within the locus. Transient expression of Cre recombinase via electroporation of pCre-AC plasmid was then used to excise the floxed PGK-driven puromycin resistance cassette, and a normal male karyotype in the resulting Nkx2-1-GFP ES cell line was confirmed by G-banding analysis. Nkx2-1GFP ES cells were injected into C57BL/6J (Jackson Laboratory) mouse blastocysts followed by implantation into pseudopregnant CD-1 foster mothers using routine methods. Germline transmission of the Nkx2-1-GFP allele was documented in the resulting chimeras by standard breeding procedures and genotyping tail snips by PCR screening using primers: 5’-AAAGTAGCGAGGCTTCGCCTTCC, 3’-CACTGCACGCCGTAGGTCAGG. Maintenance on C57BL/6J (JAX Strain #000664) background
Suggested Control Mice
C57BL/6J
MMRRC Genetic QC Summary
The MMRRC Centers have developed a genetic QC pipeline using MiniMUGA array genotyping to provide additional information on strain backgrounds for MMRRC congenic and inbred strains. For more information on when data may be available, or to request genotyping for a strain of interest, please contact csmmrrc@jax.org. Older strains may not have this information.
  • Cell Biology
  • Developmental Biology
  • Research Tools
Donor
Laertis Ikonomou, Ph.D., Boston University School of Medicine.
Darrell N. Kotton, M.D., Boston University, School of Medicine.
Primary Reference

Longmire TA, Ikonomou L, Hawkins F, Christodoulou C, Cao Y, Jean JC, Kwok LW, Mou H, Rajagopal J, Shen SS, Dowton AA, Serra M, Weiss DJ, Green MD, Snoeck HW,Ramirez MI, Kotton DN. Efficient derivation of purified lung and thyroidprogenitors from embryonic stem cells. Cell Stem Cell. 2012 Apr 6;10(4):398-411. doi: 10.1016/j.stem.2012.01.019. PubMed PMID: 22482505; PubMed Central (Medline PMCID: PMC3322392)

Colony and Husbandry Information

Cryo-recovered strains distributed by the MMRRC at JAX are shipped to the customer from the Pathogen & Opportunistic-Free Animal Room G200 - see https://www.jax.org/jax-mice-and-services/customer-support/customer-service/animal-health/health-status-reports.

Mice recovered from a cryo-archive will have health surveillance performed on recipient females. Health reports will be provided prior to shipment. If you require additional health status information, please email csmmrrc@jax.org.
Coat Color
Black
Eye
Black
MMRRC Breeding System
Backcross and sib-mating
Generation
N10 (C57BL/6J), F8
Overall Breeding Performance
Excellent
Viability and Fertility: Female Male Comments
Homozygotes are viable: No No
Homozygotes are fertile: N/A N/A
Heterozygotes are fertile: Yes Yes
Age Reproductive Decline: 7 to 9 months Greater than 12 months
Average litter size
7 to 9
Recommended wean age
4 Weeks
Average Pups Weaned
7 to 9

Order Request Information

Limited quantities of breeder mice (recovered litter) are available from a cryoarchive; recovered litter usually available to ship in 3 to 4 months.

Cryopreserved material may be available upon request, please inquire to csmmrrc@jax.org for more information.

The donor or their institution limits the distribution to non-profit institutions only.

Distribution of this strain requires submission of the MMRRC Conditions of Use (COU). A link to the COU web form will be provided via email after an order has been placed; the form should be completed then or the email forwarded to your institutional official for completion.

Additional charges may apply for any special requests. Shipping costs are in addition to the basic distribution/resuscitation fees. Information on shipping costs and any additional charges will be provided by the supplying MMRRC facility.

Click button to Request this one strain. (Use the MMRRC Catalog Search to request more than one strain.)
MMRRC Item # Description Distribution Fee / Unit (US $)
*Shipping & Handling not included*
Units Notes
066764-JAX-SPERM Cryo-preserved spermatozoa $437.00 / Non-Profit Aliquot Approximate quantity3
066764-JAX-RESUS Litter recovered from cryo-archive $2,022.00 / Non-Profit Litter Recovered litter4; additional fees for any special requests.
Cryopreserved material may be available upon request, please inquire to csmmrrc@jax.org for more information.

1 The distribution fee covers the expense of rederiving mice from a live mouse; you will receive the resulting litter. The litter will contain at minimum one mutant carrier; the actual number of animals and the gender and genotype ratios will vary. (Typically, multiple breeder pairs can be established from the recovered litter.) Prior to shipment, the MMRRC will provide information about the animals recovered. If you anticipate or find that you need to request specific genotypes, genders or quantities of mice in excess of what is likely from a resuscitated litter, you may discuss available options and pricing with the supplying MMRRC facility.

2 An aliquot contains a sufficient number of embryos (in one or more vials or straws and based on the transfer success rate of the MMRRC facility) to transfer into one to three recipients. The MMRRC makes no guarantee concerning embryo transfer success experienced in the recipient investigator's laboratory. Neither gender nor genotype ratios are guaranteed.

3 An aliquot is one straw or vial with sufficient sperm to recover at least one litter of mice, as per provided protocols, when performed at the MMRRC facility. The MMRRC makes no guarantee concerning the success of these procedures when performed outside the MMRRC facilities.

4 The distribution fee covers the expense of resuscitating mice from the cryo-archive; you will receive the resulting litter. The litter will contain at minimum one mutant carrier; the actual number of animals and the gender and genotype ratios will vary. (Typically, multiple breeder pairs can be established from the recovered litter.) Prior to shipment, the MMRRC will provide information about the animals recovered. If you anticipate or find that you need to request specific genotypes, genders or quantities of mice in excess of what is likely from a resuscitated litter, you may discuss available options and pricing with the supplying MMRRC facility.

To request material from the MMRRC: Please fill out our on-line request form (accessible from the catalog search results page, or click the Request this Strain button in the fees section). If you have questions or need assistance completing this form, you may call Customer Service at (800) 910-2291 (in USA or Canada) or (530) 757-5710 (international calls). Before you call, please have with you: the MMRRC item number, quantity needed, Bill-to and Ship-to contact information.