The SCNT (somatic cell nuclear transfer) technique is a key technology of modern biotechnology,has been widely used in livestock breeding of excellent individuals, transgenic technology and bio-pharmaceuticals.However, due to incomplete reprogramming factors, the low birth rate of cloned animals and the high proportion of abnormal individuals affect the further development of SCNT technology.Accumulating evidence suggests that some microRNAs (miRNAs) are involved in the development potential of SCNT embryos.As the abundant miRNA-101 in SCNT donor cells and bovine oocytes, numerous studies have demonstrated that miR-101 can regulate cell proliferation and apoptosis.However, its roles in bovine SCNT donor cells and embryo development are not known.In order to elucidate the roles of miR-101 in bovine SCNT donor cells and embryos, miR-101 was overexpressed in donor cells and the effects on the donor cell proliferation and embryo development were investigated.Firstly, luciferase reporter and western blot assays identified ING3 as a novel target of miR-101 in bovine fibroblasts.The results of Luciferase Reporter demonstrated that the 3-UTR in ING3 mRNA contained a complementary site for the seed region of miR-101.The protein level of ING3 was detected by western blot and the result showed that miR-101 could down-regulate ING3 protein expression.Secondly,real time PCR and flow cytometry methods were introduced to examine the effects of miR-101 on gene transcripts and cell survival.The results showed miR-101 reduced ING3 mRNA level, and also inhibited expression of P53, P21, UHRF1, BAX and DNMT3A.In addition, miR-101 promoted the growth of bovine fetal fibroblasts and decreased its apoptosis.Finally, the effects of miR-l01 overexpression on the development of SCNT embryos and blastocyst quality were evaluated.By utilizing donor cells with overexpressed miR-101, the blastocyst rate of bovine SCNT embryos in miR-101 group (35.75%) was significantly higher than the control group (26.06%) (P <0.05).And the apoptotic rate in SCNT blastocysts was reduced in miR-101 group compared to the control group (P <0.05).The genes OCT4 and SOX2 were detected higher in miR-101 group SCNT blastocysts than the control group (P <0.05).In conclusion, the present study indicates that miR-101 can reduce cell apoptosis, expedite cell cycle by targeting ING3 in bovine fibroblasts.The importantly, miR-101 can promote the development of SCNT embryos and improve embryo qualities.Therefore, miR-101 is a potential tool to enhance the nuclear reprogramming of bovine SCNT embryos.