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In a previous study, miR-2478 was demonstrated to be up-regulated in dairy goat mammary glands during peak lactation compared with the dry period. However, the detailed mechanisms by which miR-2478 regulates physiological lactation and mammary gland development in dairy goats remain unclear. In this study, we used bioinformatics analysis and homologous cloning to predict the target genes of miR-2478 and selected  INSR, FBXO11, TGFβ1  and  ING4  as candidate target genes of miR-2478. Subsequently, by targeting the 5′UTR of the  TGFβ1  gene, we verified that miR-2478 significantly inhibited  TGFβ1  transcription and the Pearson’s correlation coefficient between miR-2478 expression and  TGFβ1  expression was −0.98. Furthermore, we identified the potential promoter and transcription factor binding regions of  TGFβ1  and analyzed the potential mechanisms of interaction between miR-2478 and  TGFβ1 . Dual-luciferase reporter assays revealed that two regions, spanning from −904 to −690 bp and from −79 to +197 bp, were transcription factor binding regions of  TGFβ1 . Interesting, the miR-2478 binding sequence was determined to span from +123 to +142 bp in the  TGFβ1  gene promoter. Thus, our results have demonstrated that miR-2478 binds to the core region of the  TGFβ1  promoter and that it affects goat mammary gland development by inhibiting  TGFβ1  transcription.

miR-2478 inhibits TGFβ1 expression by targeting the transcriptional activation region downstream of the TGFβ1 promoter in dairy goats