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Meat quality and meat composition are not separated from the influences of animal genetic improvement systems; the growth and development of skeletal muscle are the primary factors in agricultural meat production and meat quality. Though the muscle-type cofilin (CFL2) gene has a crucial influence on skeletal muscle fibers and other related functions, the epigenetic modification mechanism of the <i>CFL2</i> gene regulating meat quality remains elusive. After exploring the spatiotemporal expression data of <i>CFL2</i> gene in a group of samples from fetal bovine, calf, and adult cattle, we found that the level of <i>CFL2</i> gene in muscle tissues increased obviously with cattle age, whereas DNA methylation levels of <i>CFL2</i> gene in muscle tissues decreased significantly along with cattle age by BSP and COBRA, although DNA methylation levels and mRNA expression levels basically showed an opposite trend. In cell experiments, we found that bta-miR-183 could suppress primary bovine myoblast differentiation by negatively regulated CFL2. In addition, we packaged recombinant adenovirus vectors for <i>CFL2</i> gene knockout and overexpression and found that the <i>CFL2</i> gene could promote the differentiation of primary bovine myoblasts by regulating marker genes <i>MYOD</i>, <i>MYOG</i> and <i>MYH3</i>. Therefore, CFL2 is an essential mediator for promoting myogenic differentiation by regulating myogenic marker genes in cattle myoblasts.