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The Pacific white shrimp <i>Litopenaeus vannamei</i> is the most economically important crustacean in the world. The growth and development of shrimp muscle has always been the focus of attention. Myocyte Enhancer Factor 2 (MEF2), a member of MADS transcription factor, has an essential influence on various growth and development programs, including myogenesis. In this study, based on the genome and transcriptome data of <i>L</i>. <i>vannamei</i>, the gene structure and expression profiles of <i>MEF2</i> were characterized. We found that the <i>LvMEF2</i> was widely expressed in various tissues, mainly in the Oka organ, brain, intestine, heart, and muscle. Moreover, <i>LvMEF2</i> has a large number of splice variants, and the main forms are the mutually exclusive exon and alternative 5′ splice site. The expression profiles of the <i>LvMEF2</i> splice variants varied under different conditions. Interestingly, some splice variants have tissue or developmental expression specificity. After RNA interference into <i>LvMEF2</i>, the increment in the body length and weight decreased significantly and even caused death, suggesting that <i>LvMEF2</i> can affect the growth and survival of <i>L. vannamei</i>. Transcriptome analysis showed that after <i>LvMEF2</i> was knocked down, the protein synthesis and immune-related pathways were affected, and the associated muscle protein synthesis decreased, indicating that <i>LvMEF2</i> affected muscle formation and the immune system. The results provide an important basis for future studies of the <i>MEF2</i> gene and the mechanism of muscle growth and development in shrimp.