MicroRNA-378 (miRNA-378) has been reported to have a crucial role in skeletal muscle differentiation; however, the underlying mechanisms have largely remained to be elucidated. The present study employed high‑throughput RNA sequencing to investigate the transcriptome following transfection of miRNA‑378 mimics or control RNAs into C2C12 myoblast cells. By sequencing and annotation, 2,802 transcripts that were changed by >1.5 fold were obtained and then subjected to signaling pathway enrichment and gene ontology analysis. Eight genes associated with development were subsequently selected for validation by quantitative qPCR, the results of which were highly consistent with those of the high‑throughput RNA sequencing. The protein levels of bone morphogenetic protein 4 (BMP4), which was among the differentially expressed genes, were decreased following ectopic expression of miRNA‑378. BMP4 was further confirmed to be a direct target of miRNA‑378 by using a dual luciferase assay. Finally, treatment with miRNA‑378 or small interfering RNA against BMP4 induced myogenic differentiation in C2C12 cells. In conclusion, the present study suggested that miRNA‑378 is critical for the promotion of myoblast differentiation by targeting BMP4.