BACKGROUND: The lentiviral vector is an effective tool to execute RNAi in the human dendritic cells. However, there are few reports addressing applying RNAi technology to the human dendritic cell-specific intercellular adhesion molecule 3 grabbing nonintergrin (DC-SIGN).
OBJECTIVE: To construct a lentiviral vector expressing small-hairpin RNA (shRNA) targeting human DC-SIGN gene, and to provide a useful tool to regulate the expression level of DC-SIGN.
METHODS: Four target sequences were selected according to human DC-SIGN mRNA sequence．The cDNA target sequence containing both sense and antisense Oligo DNA were designed. The obtained lentiviral vector containing DC-SIGN shRNA was confirmed by PCR, sequencing and exogenous selection. 293T cells were cotransfected with lentiviral vector pGCSIL-GFP, pHelper1.0 and pHelper 2.0. The titer of virus was tested by serial dilution．
RESULTS AND CONCLUSION: PCR and DNA sequencing demonstrated that the inserted sequences were correct. Two effective targeting sequences were determined by exogenous selection. The titer of concentrated virus was 1×10⁹ T U/mL. The lentivirus vector targeting DC-SIGN has been successfully constructed.