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Nkx2.5-iCre Mouse Model
| Strain Name | C57BL/6NMcl- Nkx2.5<sup>tm1(IRES-NLS-iCre)</sup>/MCL |
|---|---|
| Editing Type | Knock-In |
| Gene Name | Nkx2-5/NK2 homeobox 5 |
| Gene Synonyms | Csx, Nkx2.5, Tinman |
| Strain Background | C57BL/6N |
| NCBI No. | Gene ID: 18091 / MGI: 97350 |
- Cardiac progenitor cell fate determination and lineage tracing
- Studies on embryonic cardiogenesis and heart tube formation mechanisms
- Research on the regulation of cardiomyocyte and endocardial cell differentiation
- Research on the developmental mechanisms of the cardiac outflow tract, atrioventricular canal, and septa
- Functional studies of genes associated with congenital heart disease
- Research on the mechanisms of cardiomyopathy, cardiac conduction abnormalities, and cardiac remodeling
- Constitutive conditional knockout, knock-in, or reporter activation of floxed genes related to cardiac development
By precisely inserting an IRES-NLS-iCre expression cassette into the 3′ untranslated region (3′UTR) immediately downstream of the Nkx2-5 stop codon, the Nkx2-5-IRES-NLS-iCre knock-in allele was successfully generated. In this model, Nkx2-5 and the downstream NLS-iCre are co-transcribed from a single transcript; the IRES element mediates independent translation of the downstream NLS-iCre open reading frame, such that the NKX2-5 transcription factor and the NLS-iCre recombinase are expressed as separate proteins. The NLS sequence promotes nuclear localization of iCre, enabling recognition of loxP sites flanking floxed alleles and mediating DNA recombination. iCre is a mammalian codon-optimized improved Cre recombinase that enhances its expression and loxP recombination efficiency in mouse cells [1,2].
The Nkx2-5 gene encodes the NK2 homeobox transcription factor NKX2-5, a critical regulator of embryonic cardiogenesis. Nkx2-5 is expressed in the cardiac crescent, heart tube, and cardiac progenitors, and participates in cardiomyocyte differentiation, cardiac morphogenesis, and transcriptional regulation of numerous cardiac developmental genes [3]. Previous studies have demonstrated that the Nkx2-5 3′UTR-IRES-Cre knock-in strategy achieves efficient recombination at the cardiac crescent and heart tube stages, and labels Nkx2-5-positive cardiac progenitors and their derivatives, including myocardium, endocardium, dorsal mesocardium, and pericardial mesenchyme [1,4]. Therefore, when Nkx2-5-IRES-NLS-iCre mice are crossed with mice carrying floxed alleles, constitutive conditional gene knockout, knock-in, or lineage tracing can be achieved in Nkx2-5-positive cells and their progeny, providing a genetic tool for studying cardiac progenitor cell fate determination, myocardial and endocardial development, outflow tract formation, cardiac structural morphogenesis, and the mechanisms of congenital heart disease.
References
[1] Stanley EG, Biben C, Elefanty A, Barnett L, Koentgen F, Robb L, et al. Efficient Cre-mediated deletion in cardiac progenitor cells conferred by a 3′UTR-IRES-Cre allele of the homeobox gene Nkx2-5. Int J Dev Biol. 2002;46(4):431-439.
[2] Shimshek DR, Kim J, Hübner MR, Spergel DJ, Buchholz F, Casanova E, et al. Codon-improved Cre recombinase (iCre) expression in the mouse. Genesis. 2002;32(1):19-26.
[3] Tanaka M, Chen Z, Bartunkova S, Yamasaki N, Izumo S. The cardiac homeobox gene Csx/Nkx2.5 lies genetically upstream of multiple genes essential for heart development. Development. 1999;126(6):1269-1280.
[4] Zhou B, von Gise A, Ma Q, Rivera-Feliciano J, Pu WT. Nkx2-5- and Isl1-expressing cardiac progenitors contribute to proepicardium. Biochem Biophys Res Commun. 2008;375(3):450-453.
- Endogenous Expression Drive: NLS-iCre expression is controlled by endogenous Nkx2-5 regulatory elements, coupling the recombination domain to the native Nkx2-5 expression pattern.
- Independent Protein Expression: The IRES element mediates separate translation of NKX2-5 and NLS-iCre, allowing both proteins to function independently.
- Efficient Nuclear Recombination: The NLS promotes iCre nuclear import, enabling loxP recombination in Nkx2-5-positive cells.
- Early Embryonic Onset: Recombination initiates at the cardiac crescent and heart tube stages, ideal for early cardiac development studies.
- Cardiac Progenitor Coverage: Labels Nkx2-5-positive cardiac progenitors and their derivatives, including myocardium and endocardium.
FAQ
Game-changing benefits？
While competitors highlight germline efficiency gains, shorter timelines and enhanced 3Rs animal welfare benefits for their technologies, these are merely incremental improvements over traditional approaches. In sharp contrast, our proprietary technology delivers fully pure, homogeneous lineages—every single cell of the mice is derived exclusively from totipotent ES cells, with guaranteed 100% germline transmission efficiency. To experience these unparalleled benefits firsthand, enquire about your custom mouse model project with us or order embryos for in-house validation at your facility.
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