Acta Veterinaria et Zootechnica Sinica ›› 2024, Vol. 55 ›› Issue (3): 984-994.doi: 10.11843/j.issn.0366-6964.2024.03.012

• ANIMAL GENETICS AND BREEDING • Previous Articles     Next Articles

Regulation of NO Concentration by NOS2 Gene DNA Methylation Editing Affects the Expression of Muscle Development Pathway Genes

SHEN Qi1,2,3, WANG Kai4, ZHAO Zhenjian1,2,3, CHEN Dong1,2,3, YU Yang1,2,3, CUI Shengdi1,2,3, WANG Junge1,2,3, CHEN Ziyang1,2,3, WU Pingxian5, TANG Guoqing1,2,3*   

  1. 1. Key Laboratory of Livestock and Poultry Multi-omics of Ministry of Agriculture and Rural Affairs, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, China;
    2. Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu 611130, China;
    3. State Key Laboratory of Swine and Poultry Breeding Industry, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, China;
    4. New Hope Liuhe Co. Ltd., Beijing 100102, China;
    5. National Center of Technology Innovation for Pigs, Chongqing 402460, China
  • Received:2023-09-01 Online:2024-03-23 Published:2024-03-27

Abstract: This study aimed to explore the regulation mechanism of DNA methylation editing of nitric oxide synthase (NOS) 2 on Myoglobin, and study its effect on the expression level of neuronal nitric oxide synthase (nNOS) and endothelial nitric oxide synthase (eNOS). In this experiment, DNA methylation editing technology was used to methylate NOS2 in porcine kidney cell line PK-15 cells, and the DNA methylation level of the NOS2 gene promoter, the expression level of NOS2, the concentration of intracellular nitric oxide, as well as the expression level of myofibrillar-and myoglobin-related genes were detected in porcine kidney cells. The results revealed that the average DNA methylation level of the first 500 bp region of the NOS2 gene promoter was reduced, but the difference was not significant, while the methylation level of the CpG site near the gRNA2 binding site was increased. In addition, after DNA methylation editing of NOS2 gene, NOS2 gene expression and its iNOS protein expression were significantly reduced. With the decrease of NOS2 expression, nNOS and eNOS expression increased significantly, the expression level of MYHC-Ⅱb and MYHC-Ⅱx decreased significantly, and the expression of Myoglobin protein decreased significantly. In conclusion, the abnormal expression of the NOS2 gene can affect the generation of NO in cells, as well as the expression of NOS1 and NOS3, and the expression of genes related to muscle development.

Key words: NOS2, DNA methylation, NO, muscle fiber, myoglobin

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