Lentivirus-Mediated RNA Interference of SOCS3 Gene Expression in Porcine Preadipocytes and Myoblasts

This study is to construct lentivirus interfering vector target on porcine suppressor of cytokine signaling 3,obtain infectious lentivirus particles,and observe its interference efficiency of porcine preadipocyte.Three pairs of small interfering RNA(siRNA) target SOCS3 cDNA were designed,synthesized and linked with lentivirus vector LentiH1 to construct the lentivirus recombinant plasmids.After sequencing confirmation,each of lentivirus recombinant plasmide and packaging plasmids were cotransfected into 293T cells.The recombinant lentivirus were packaged in 293T cells.After purification,virus titer were determined.Restriction enzyme and DNA sequencing demonstrated that the recombinant lentivirus vector was constructed correctly,and the virus titer reached 3×107 tu /mL,RT-PCR and Western blotting showed that SOCS3 mRNA and protein expression were remarkably decreased in porcine preadipocytes infected with recombinant lentivirus,of which,SOCS3 mRNA and protein expression were suppressed by approximate 53% and 71% by LentiH1-siRNA3,respectively.In conclusion,this study successfully constructed the recombinant lentivirus interfering vector target on porcine SOCS3 gene,it may lay a solid foundation for the further research on the function of SOCS3.