To construct one lentiviral vector containing mouse SRY-related silencing group--box gene 9 (SOX9) and to transfect murine bone mesenehymal stem cells (mBMSCs) in vitro and observe the expression of target gene.RNA inteference target sequence was designed in connectin with mice SOX9 gene sequence. The double strands DNAoligo containing interference sequence were synthesized and cloned into lentivirus vector. The siRNA lentiviral vector with SOX9 gene silencing was constructed and identified, which was transfected into rat bone mesenehymal stem cells. The expression of target gene was detected by immunofluorescence, RT-PCR and Western blot.Lenti-SOX9-siRNA-EGFP was recombined successfully and transduced efficiently into mBMSCs. The expression of SOX9 gene silencing was confirmed by RT-PCR and Western blot.Mouse SOX9 gene silencing by RNA interference and Lentiviral vector can transfected successfully into mBMSCs. Meanwhile,SOX9 gene may be silenced in SOX9 transduced mBMSCs. This will provide target cells for the following study about SOX9 gene respairing cartilage injury.