Use of Isl1 gene or protein in Tetralogy of Fallot (TOF) diagnosis
A Tetralogy of Fallot, gene technology, applied in the direction of biochemical equipment and methods, microbe measurement/testing, material inspection products, etc., can solve problems such as cardiac hypoplasia
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Embodiment 1
[0099] Example 1 Detailed phenotypic analysis of Isl1 mutant mice to elucidate the role of Isl1 in the formation of cardiac structures
[0100] method:
[0101] (1) Animal model
[0102] Mating Isl1hypo / hypo with Isl1+ / nLacZ or Isl1hypo / hypo will generate Isl1 compound low mutant mice (Isl1hypo / nLacZ) and low expression mutant mice, respectively. TnT-Cre;Isl1 f / + Mating with Isl1f / f mice will generate cardiomyocyte-specific Isl1 knockout mouse variants. Embryos were collected at different time points.
[0103] (2) Histological analysis: Mice were crossed, embryos were harvested at the desired developmental stages, and fixed with 4% PFA for whole-body and section staining analysis. For high-resolution tissue analysis, whole-mount embryos were paraffin-embedded and sectioned, and stained with Nuclear Fast Red. To analyze the morphology of the pharyngeal arch arteries, both transverse and longitudinal sections will be analyzed.
[0104] (3) Immunostaining and Cell Prolifera...
Embodiment 2
[0107] Example 2 Molecular Mechanism of Isl1 in the Process of Cardiac Outflow Tract Formation
[0108] (1) RNA in situ hybridization. Using Digoxigenin-labeled cRNA probe and AP-labeled Digoxigenin antibody (Roche Company), BCIP and NBT (Roche Company) kits were used for color development. The probes used include: BMP, TGF, FGF, Wnts, Shh, Notch, endothelins, PDGF, Tbx1, Tbx3 / 5, GATA factors, Nkx2.5, Mef2c, Foxh1, etc.
[0109] (2) Gene chip analysis: According to the type of change in outflow tract vascular phenotype and the time point of appearance, the time point of collection of outflow tract specimens will be determined, and the control, TnT-Cre; Isl1 variant and Isl1 low expression variation will be collected Mouse outflow tract specimens. The cardiac outflow tract samples were immediately placed in RNAlater and kept overnight at 4°C, and the RNAlater was removed the next day and stored at -80°C. RNA was extracted with Trizol. cRNA synthesis, labeling, microarray hy...
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