Single-nucleotide Resolution Epitranscriptomic Profiling Uncovers Dynamic m 6 A Regulation in Bovine Preimplantation Development

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Abstract RNA N 6 -methyladenosine (m 6 A) plays a crucial role in regulating gene expression during early embryonic development. However, the m 6 A dynamics at single-nucleotide resolution in preimplantation development remain uncharacterized, and the functional significance of site specific m 6 A modifications in key developmental regulators is largely unknown. Here, using SAC-seq, a single-base resolution, antibody-independent m 6 A profiling method, we generate the first comprehensive m 6 A landscape in bovine oocytes and preimplantation embryos. We identify a previously uncharacterized m 6 A site in RPL12 transcript that is essential for embryonic development. Loss of m 6 A at this site leads to reduced protein synthesis, disrupted expression of translation-related genes, and impaired zygotic genome activation and blastocyst formation. Notably, supplementation with wild-type RPL12 mRNA fails to rescue the developmental arrest, indicating that m 6 A regulation extends beyond transcript abundance. Our findings provide a valuable resource of m 6 A at single-nucleotide resolution in mammalian embryogenesis and uncover a critical mechanism by which precise, site-specific m 6 A regulates translation and developmental competence in early embryos.

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