PMID: 11933217Apr 5, 2002Paper

High efficiency lentiviral gene delivery in non-dividing cells by deoxynucleoside treatment

The Journal of Gene Medicine
Elisabetta RavotJulianna Lisziewicz

Abstract

Gene therapy has recently been advanced by the development of HIV-based vectors that are able to transduce some non-dividing cells. The manipulation of most non-dividing cells remains, however, scarcely efficient. One of the biological mechanisms postulated to prevent powerful transduction of quiescent cells by lentiviral vectors is the paucity of deoxynucleotides (dNTPs). In this study, a novel delivery strategy is developed to improve significantly the efficiency of HIV-based vectors in transducing non-dividing cells. This approach is based on increasing the intracellular availability of dNTPs by incubating target cells with the dNTP precursors, deoxynucleosides (dNSs). Mature human monocyte-derived macrophages (14-21 days old) were transduced at a low multiplicity of infection (MOI) of HIV vectors carrying a reporter gene. dNSs were added to the medium during transduction (5 mM dNS) and immediately before post-transduction culture (2.5 mM dNS). Macrophages were harvested 2-7 days after transduction and assayed for transgene expression by cytofluorimetry. The addition of dNS to the medium significantly enhanced the efficiency of transduction of human macrophages by HIV-based vectors. The percentage of cells expressing the tra...Continue Reading

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Citations

Jan 21, 2010·Molecular Therapy : the Journal of the American Society of Gene Therapy·Janka MátraiThierry VandenDriessche
Mar 25, 2005·The Journal of Biological Chemistry·Pritam SenguptaBarbara D Smith
Aug 29, 2012·Nature Reviews. Microbiology·Diana AyindeOlivier Schwartz

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