Overexpression of miR-320-3p, miR-381-3p, and miR-27a-3p Suppresses Genes Related to Midline Facial Cleft in Mouse Cranial Neural Crest Cells

Midline facial clefts are severe craniofacial defects that occur due to an underdeveloped frontonasal process. While genetic studies in mice have identified several genes that are crucial for midfacial development, the interactions and regulatory mechanisms of these genes during development remain unclear. In this study, we conducted a systematic review and database search to curate genes associated with midline facial clefts in mice. We identified a total of 78 relevant genes, which included 69 single-gene mutant mice, nine spontaneous models, and 20 compound mutant mice. We then performed bioinformatic analyses with these genes to identify candidate microRNAs (miRNAs) that may regulate the expression of genes related to midline facial clefts. Furthermore, we experimentally evaluated the four highest-ranking candidates-miR-320-3p, miR-381-3p, miR-27a-3p, and miR-124-3p-in O9-1 cells. Our results indicated that overexpression of any of these miRNAs inhibited cell proliferation through the suppression of genes associated with midline facial clefts. Thus, our results suggest that miR-320-3p, miR-381-3p, miR-27a-3p, and miR-124-3p are involved in the cause of midline facial anomalies.