Establishment of transgenic mouse model with cardiac overexpression of PRKAG2-G100S / 第二军医大学学报

ABSTRACT

Objective To establish a novel transgenic mouse model of human PRKAG2 cardiac syndrome that overexpresses a PRKAG2G100S mutation, so as to lay a foundation for further studying the role of human PRKAG2 gene in the development, morphology, and function of mouse heart. Methods Human PRKAG2 with G100S mutation was sub-cloned into a multiple cloning site located in the downstream of α-myosin heavy chain (a-MHC) promoter of the plasmid. After the construction of the transgenic expressing vector, C57BL/6J mice were selected as the genetic background, and the transgenic mouse model of PRKAG2-G100S mutation was buitt by microinjection. Genotype was further confirmed using specific primer PCR. Real time PCR and Western blotting analysis were used to examin the expression of human PAKAG2(G100S) mRNA and protein, respectively. Results Two strains of transgenic mice were successfully developed using backcross breeding, which specifically overexpressed the human PRKAG2-G100S mutation in the cardiac tissues of F2 generations by the methods qPCR and Western blotting at both mRNA and protein levels. Moreover, the PRKAG2-G100S mutation was successfully passed steadily. Conclusion We have successfully established a human PRKAG2-G100S transgenic mouse model, which can help to further explore the role of PRKAG2-G100S mutation in the development and function of mouse cardiac tissue in the PRKAG2- G100S cardiac syndrome.