Objective       To construct and identify miRNA expression vectors interfering mouse core binding factor β subunit (Cbfβ) gene and to determine their effects on osteogenic differentiation of mesenchymal stem cell C3H10T1/2.       Methods       Artificial microRNA sequences specifically interfering mouse Cbfβ gene were designed and Blockit miRNA-Cbf-β-miRNA1/miRNA2 vectors interfering mouse Cbfβ gene were constructed. The vectors were transferred into C3H10T1/2 cells to get stable cell line. Quantitative RT-PCR and Western blotting were used to detect the expression levels of Cbfβ

Runt-related tra

nscription factor 2 (Runx2) and osteogenesis marker gene alkaline phosphatase (ALP) in the cells in 3 d after BMP-2 treatment.       Results       Artificial microRNA vectors interfering Cbfβ were successfully constructed and stable cell line was obtained. The expression level of ALP and Cbfβ were obviously reduced （

P

0.05）

while Runx2 expression was not notably decreased （

P

0.05）.       Conclusion       miRNA expression vectors miRNA-Cbf-β-miRNA1/miRNA2 targeting Cbfβ gene may inhibit function of osteogenic transcription factor Cbfβ and then decrease protein and mRNA expression of ALP. Stable cell line with influenced osteogenic ability is obtained.