Abstract:
:Lentiviral vectors derived from the human immunodeficiency type 1 virus (HIV-1 LV) are among the finest tools available today for the genetic modification of human monocyte-derived dendritic cells (MDDCs). However, this process is largely inefficient because MDDCs show a strong resistance to HIV-1 transduction. Here we describe a step-by-step protocol from the production of LVs to cell transduction that allows the efficient genetic modification of MDDCs. This protocol can be completed in 23 d from the initial phase of LV production to the final analysis of the results of MDDC transduction. The method relies on the simultaneous addition of HIV-1 LVs along with noninfectious virion-like particles carrying Vpx, a nonstructural protein encoded by the simian immunodeficiency virus (Vpx-VLPs). When thus provided in target cells, Vpx exerts a strong positive effect on incoming LVs by counteracting the restriction present in MDDCs; accordingly, 100% of cells can be transduced with low viral inputs. Vpx-VLPs will improve the efficiency of LV-mediated transduction of MDDCs with vectors for both ectopic gene expression and depletion studies.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Berger G,Durand S,Goujon C,Nguyen XN,Cordeil S,Darlix JL,Cimarelli Adoi
10.1038/nprot.2011.327subject
Has Abstractpub_date
2011-06-01 00:00:00pages
806-16issue
6eissn
1754-2189issn
1750-2799pii
nprot.2011.327journal_volume
6pub_type
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