Gene Transduction in Human Monocyte-Derived Dendritic Cells Using Lentiviral Vectors
Monocyte-derived dendritic cells (MDDCs) are widely used in the field of human immunology. Although a variety of gene delivery procedures have been used in MDDC, it has remained difficult to achieve robust gene transductions. In this chapter, we describe a procedure for high efficiency gene transduction in human MDDCs using lentiviral vectors. Gene transduction based on HIV-1-derived lentiviral vectors is restricted at the level of reverse transcription by the cellular protein SAMHD1 in MDDCs. Co-transduction of the MDDCs with helper particles derived from SIVmac that contain the viral protein Vpx removes this restriction, leading to a drastic increase in the rate of gene transduction. This procedure leads to nontoxic, efficient and stable transduction in MDDCs. It can be applied to any HIV-1-derived lentiviral vector, including shRNA lentiviral vectors for RNAi. Transduced MDDCs are not activated by the transduction and can be activated normally by TLR ligands.
Source: Springer protocols feed by Immunology - Category: Allergy & Immunology Source Type: news
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