Abstract
The rhesus macaque is one of the closest evolutionary relatives to humans, making the study of alternative splicing (AS) during its early embryonic development highly valuable for understanding human embryogenesis and related diseases. However, systematic studies in this context remain limited. Here, a comprehensive bioinformatic analysis of AS was performed using RNA-seq data spanning early rhesus macaque embryogenesis. We identified multiple previously unannotated zygotic genome activation (ZGA) genes, thereby refining the rhesus macaque ZGA gene repertoire. The landscape of AS and differential AS events (DASEs) across early stages was characterized, revealing dynamic and stage-specific regulation, with a marked increase in AS events from the 8-cell to morula stages. In addition, weighted gene co-expression network analysis identified 35 key splicing factors (SFs) involved in regulating early rhesus macaque embryonic development. Finally, we calculated the correlation between differentially expressed SFs and DASEs during the ZGA process, and identified potential regulatory relationships between several SFs (TRA2B, IGF2BP1, HNRNPAB, and MATR3) and specific DASEs. Collectively, this study provides the first systematic analysis of AS dynamics and regulation in early rhesus macaque embryogenesis, highlighting its critical role in development and offering a valuable reference for understanding AS in early human embryos.