Abstract Nutrients are utilized and re-constructed by endodermal epithelial cells (EECs) in yolk sac membranes in avian species. Sterol-O acyltransferase 1 (SOAT1) is the key enzyme to convert cholesterol to cholesteryl ester for delivery to growing embryos. During development, absorption of yolk is matched with significant changes of SOAT1 mRNA and enzyme activity. miRNAs regulate angiogenesis and metabolism during mammalian development. However, the involvement of miRNAs in lipid utilization during avian embryogenesis remains ambiguous. Using a miRNA sequencing technique, we found several candidate miRNAs and confirmed expression patterns with real time PCR. They were selected for as candidates targeting the receptor (TGFβ receptor type 1, TGFBR1) that may regulate SOAT1. Similar to SOAT1 mRNA accumulation, the gga-miR-181a-5p expression was gradually elevated during development, but the concentration of gga-miR-429-3p was in the opposite direction. Transfection with gga-miR-181a-5p or gga-miR-429-3p inhibited TGFBR1 and SOAT1 in EECs. The 3’ untranslated region (3’UTR) of TGFBR1 was then confirmed to be one of the targets of gga-miR-181a-5p and gga-miR-429-3p. Taken together, expression of miRNAs during embryonic development regulates SOAT1 expression by inhibiting the 3’UTR of TGFBR1. This is indicative of possible regulation of avian yolk lipid utilization and modification of hatchability by changing miRNA expressions.