These Fgd5-CreERT2 knock-in/knock-out mice express ZsGreen and tamoxifen inducible Cre recombinase from the endogenous promoter/enhancer elements of the Fgd5 locus. When induced, cre activity is observed in hematopoietic stem cells and endothelial cells. This strain also serves as a reporter strain, and may be suitable for identification and purification of hematopoietic stem cells based on single-color fluorescence.
Derrick J. Rossi, Harvard University/Boston Children’s Hospital
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
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Targeted (Recombinase-expressing, Reporter, Inducible) | Fgd5 | FYVE, RhoGEF and PH domain containing 5 |
The Fgd5 gene encodes a protein that upregulates CDC42 activity and is a marker for
immunophenotypic hematopoietic stem cells. These Fgd5ZsGr•CreERT2 mice carry a knock-in/knock-out mutation of the Fgd5 gene in which ZsGreen and creERT2 cassettes (separated
by a 2A peptide cassette) have been inserted in frame into the first coding exon. Under control of the endogenous upstream promoter/enhancer elements, tamoxifen-inducible cre activity is observed in ZsGreen+ LSKCD150+ hematopoietic stem cells. Homozygotes are not viable, homozygous embryos die around E11.5–E12.0.
The Cre-ERT2 fusion protein consists of Cre recombinase fused to a triple mutant form of the human estrogen receptor which does not bind its natural ligand (17β-estradiol) at physiological concentrations but will bind the synthetic estrogen receptor ligands 4-hydroxytamoxifen (OHT or tamoxifen) and, with lesser sensitivity, ICI 182780. Restricted to the cytoplasm, Cre-ERT2 can only gain access to the nuclear compartment after exposure to tamoxifen. To counteract the mixed estrogen agonist effects of tamoxifen injections, which can result in late fetal abortions in pregnant mice, progesterone may be coadministered.
A targeting vector was used to insert ZsGreen and creERT2 cassettes, separated
by a 2A peptide cassette, in frame into exon 1 with a FRT site flanked PGK-NEO cassette downstream of the ZsGreen cassettes.
The construct was electroporated into C57BL/6NTac derived B6-3 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were tested for germline transmission. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6NJ (Stock No. 005304) at least once to establish the colony.
Expressed Gene | GFP, Green Fluorescent Protein, |
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Expressed Gene | cre/ERT2, Cre recombinase and estrogen receptor 1 (human) fusion gene, |
Site of Expression | When induced, cre activity is observed in hematopoietic stem cells and endothelial cells. |
Allele Name | targeted mutation 3, Derrick Rossi |
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Allele Type | Targeted (Recombinase-expressing, Reporter, Inducible) |
Allele Synonym(s) | Fgd5ZsGr*CreERT2 |
Gene Symbol and Name | Fgd5, FYVE, RhoGEF and PH domain containing 5 |
Gene Synonym(s) | |
Expressed Gene | GFP, Green Fluorescent Protein, |
Expressed Gene | cre/ERT2, Cre recombinase and estrogen receptor 1 (human) fusion gene, |
Site of Expression | When induced, cre activity is observed in hematopoietic stem cells and endothelial cells. |
Strain of Origin | C57BL/6NTac |
Chromosome | 6 |
Molecular Note | The targeting vector consists of ZsGreen and creERT2 cassettes separatedby a 2A peptide cassette and followed by an FRT-flanked neomycin cassette inserted in frame into exon 1. |
When maintaining a live colony, heterozygous mice may be bred together, to wildtype siblings, or to C57BL/6NJ inbred mice (Stock No. 005304). Homozygotes are not viable.
When using the Fgd5ZsGr•CreERT2 mouse strain in a publication, please cite the originating article(s) and include JAX stock #027789 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous or wildtype for Fgd5<tm3(cre/ERT2)Djr> |
Frozen Mouse Embryo | C57BL/6N-Fgd5<tm3(cre/ERT2)Djr>/J | $2595.00 |
Frozen Mouse Embryo | C57BL/6N-Fgd5<tm3(cre/ERT2)Djr>/J | $2595.00 |
Frozen Mouse Embryo | C57BL/6N-Fgd5<tm3(cre/ERT2)Djr>/J | $3373.50 |
Frozen Mouse Embryo | C57BL/6N-Fgd5<tm3(cre/ERT2)Djr>/J | $3373.50 |
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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. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.
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