The MyoD1 Promoted Muscle Differentiation and Generation by Activating CCND2 in Guanling Cattle

SIMPLE SUMMARY: Muscle proliferation and differentiation is a complex process, which is mainly regulated by the myogenic regulatory factors (MRFs) gene family. As one of its family members, myogenic differentiation 1 (MyoD1) is an important regulator of myoblast differentiation and fibrogenesis, and its main function is to regulate the proliferation of myoblasts and the directed development of myogenic cell lines. In this study, CRISPR/CAS9 technology was used to construct the MyoD1 gene knockout MDBK cell lines, and then the transcriptome of MDBK cells was analyzed based on high-throughput sequencing technology. The differentially expressed genes in the transcriptome and the functional enrichment of differentially expressed genes were analyzed. The differentially expressed genes in the transcriptome and the functional enrichment were analyzed to provide experimental data for further studies on the mechanism of MyoD1 regulating CCND2 in myocyte differentiation. ABSTRACT: The purpose of this study was to analyze the transcriptome of MyoD1 gene knockout MDBK cells (bovine kidney cells) using high-throughput sequencing. For the first time, CRISPR/CAS9 technology was used to construct a MyoD1 knockout in MDBK cells and transcriptome sequence analysis was used to examine MyoD1-related target gene expression. Transcriptome sequencing indicated the presence of 723 differentially expressed genes (DEGs) by comparing wild type and MyoD1 knockout MDBK cells and included 178 upregulated and 72 downregulated genes. The DEGs are mainly enriched in Pl-3-kinase and AKT, p53 signaling pathways. Quantitative RT-PCR confirmed that PDE1B, ADAMTS1, DPT, and CCND2 were highly expressed in the leg muscle, longissimus dorsi, and shoulder of Guanling cattle, and CCND2 was inhibited after MyoD1 knockout, suggesting it may be a key downstream gene of MyoD1 and associated with muscle formation and differentiation in Guanling cattle. This provides experimental data for subsequent studies on the regulatory mechanisms of muscle differentiation in Guanling cattle.