Establishment and Identification of Cardiomyocyte-specific High Expression Human APE1 Transgenic Mouse

Abstract

The aim of this study is to establish transgenic mice that specifically over-express human APE1 incardiac, and to provide a tool animal for studying the relationship between the function and mutation of hAPE1 gene and cardiac development and cardiovascular diseases. A cardiomyocyte-specific hAPE1 transgenic construct containing the α-myosin heavy chain (α-MHC) promoter and human A PE1 (hA PE1) gene was generated. The transgenic vector was constructed by insertion of hAPE1 gene under the α-MHC promoter. The transgenic mice were generated by fertilized egg microinjection followed by embryo transplantation and were all maintained on C57BL/6J genetic background. The genotype of transgenic mice was identified using PCR and the expression levels of hAPE1 in different tissues were detected by Western blotting. The results indicated that cardiomyocyte-specific hAPE1 transgenic construct containing the α-myosin heavy chain (α-MHC) promoter and human APE1 (hAPE1) gene were introduced into fertilized zygotes by microinjection, and then the fertilized zygotes were implanted into the oviduct of female mice, establishing a heart specific high expression hAPE1 transgenic mouse line. 40 offspring were obtained and 15 mice carrying the human APE1 gene was identified by PCR. The heart-specific overexpression of hAPE1 was confirmed by Western blotting assay. The present study successfully obtained cardiomyocyte-specific hAPE1 transgenic mice transgenic mice expressing, which provided a useful tool for studying the function of genes in heart development and cardiovascular disease. and the transgenic mice with cardiac specific overexpression of hAPE1 were obtained. Real-time quantitative PCR and Western blotting were used to detect the expression of mRNA and protein in hAPE1 in the heart of transgenic mice. It was found that the expression level of mRNA and protein in hAPE1 in the heart of transgenic mice increased significantly. Our research showed that we have successfully constructed α - MHC-hAPE1 transgenic mice, which would provide an ideal experimental animal model for studying the detailed mechanism of hAPE1 gene in heart development and cardiovascular disease.