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Citations
IMMUNITY 47:107 (2017) IF=22.845
Tnni3-Cre Mice
Catalog Number: C001562
Strain Name: C57BL/6JCya-Tnni3em1(Cre)/Cya
Genetic Background: C57BL/6JCya
Reproduction: Carrier x WT
Examples of Expressing Tissues/Cells: Cardiomyocytes
Strain Description
Troponin I (TnI), along with Troponin T (TnT) and Troponin C (TnC), forms the troponin complex in striated muscle. TnI, the inhibitory subunit, blocks actin-myosin interactions, facilitating muscle relaxation and providing a calcium-sensitive switch for muscle contraction. The TnI subfamily includes three genes, with TNNI3 encoding cardiac-specific TnI, expressed only in heart muscle [1]. Mutations in TNNI3 cause familial hypertrophic cardiomyopathy type 7 (CMH7) and restrictive cardiomyopathy (RCM). TnI helps diagnose heart failure and ischemic heart disease, and elevated troponin levels indicate acute myocardial injury in moderate/severe COVID-19 patients. In COVID-19-related cardiovascular diseases, elevated troponin correlates with higher mortality risk.
Tnni3-Cre mice were constructed by integrating the Cre-rBG pA gene expression component into the mouse Tnni3 gene by gene editing, a strategy that disrupts the expression of the endogenous Tnni3 gene in mice. When Tnni3-Cre mice are crossed with mice containing loxP sites, sequence recombination mediated by Cre recombinase occurs in the offspring's cardiomyocytes.
Strain Strategy
The Cre-rBG pA cassette was inserted 2 bp upstream of the ATG start codon in the mouse Tnni3 gene, disrupting the expression of the endogenous Tnni3 gene.
Validation Data
a. Method
Tnni3-Cre mice were crossed with Rosa26-LSL-tdTomato mice, which conditionally express a red fluorescent protein (tdTomato). Cre recombinase-mediated deletion of the stop element (LSL) will result in tdTomato protein expression in Cre-positive cells. At 10 weeks of age, collect cardiac tissue from the offspring, and observe tissue autofluorescence using frozen section staining. Collect lung, pancreas, spinal cord, brain, ovary, and testis tissues from the offspring, and analyze tdTomato protein expression using immunofluorescence (IF) staining to determine Cre recombinase expression.
b. Group
Cre+: Tnni3-Cre[KI/+];Rosa26-LSL-tdTomato[CKI/+];
Cre-: Rosa26-LSL-tdTomato[CKI/+].
c. Result
- 1) Expression of Cre recombinase in the heart
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Figure 1. Autofluorescence of tdTomato protein in cardiac tissue. Histological morphological analysis shows a large amount of orange autofluorescence signal observed in the cardiomyocytes of Cre+ mice, indicating Cre recombination in these cells. No significant fluorescence signal was detected in the corresponding tissue of the control group (Cre-) mice.
- 2) Expression of Cre recombinase in the lung, pancreas, and spinal cord
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Figure 2. Immunofluorescence (IF) detection of tdTomato protein in lung, pancreas, and spinal cord. Results show partial red fluorescence signals in the lungs, pancreas, and spinal cord of Cre+ mice (with a non-positive red background signal), indicating Cre recombination in these tissues.
- 3) Expression of Cre recombinase in ovary and testis
Figure 3. Immunofluorescence (IF) detection of tdTomato protein in ovaries and testes. Histological morphological analysis shows partial red fluorescence signals in the luteal cells and ovarian stroma of Cre+ mice, indicating Cre recombination in these tissues. No significant fluorescence signal was observed in the testes, indicating no Cre recombination in these tissues.- d. Summary
In the Tnni3-Cre mouse model, Cre recombinase is significantly expressed in the cardiomyocytes of adult mice. Additionally, fluorescent recombination signals are detected in the lungs, pancreas, spinal cord, and ovaries (luteal cells, ovarian stroma cells) of the offspring, while no fluorescent recombination signals are observed in the testes. -
Announcements -
The Cre recombinase gene expression cassette is integrated into chromosome 7 in this strain. It is recommended that breeding with Flox mice targeted on the same chromosome is avoided.
References
[1] Bhavsar PK, Brand NJ, Yacoub MH, Barton PJ. Isolation and characterization of the human cardiac troponin I gene (TNNI3). Genomics. 1996 Jul 1;35(1):11-23.
