Construction and suppressive effect of siRNA expression vector targeting mouse 4-1BB in vitro

Objective To construct the small interfering RNA(siRNA) expression vector specific to mouse 4-1BB gene and observe its silencing effect in vitro and identify the one with high efficiency.Methods The pENTRTM/U6 siRNA expression vectors were constructed by gene recombination,then transfected into the cultured COS-7 cells.At 48h after transfection,the mRNA and protein level were separately detected by RT-PCR and FACS.Results The pENTRTM/U6 siRNA expression vectors were successfully constructed.And LipofectamineTM 2000 mediated gene transfection of p498 siRNA expression vector into COS-7 cells downregulated the mRNA and protein level of 4-1BB at 48h after transfection.Conclusion The pENTRTM/U6-p498 siRNA expression vector can specifically and efficiently inhibit 4-1BB gene expression,which may provide a foundation to construct 4-1BB Lentivirus and study 4-1BB gene function in mouse T lymphocytes.