Abstract
BackgroundLentiviral vectors have shown immense promise as vehicles for gene delivery to non-dividing cells particularly to cells of the central nervous system (CNS). Improvements in the biosafety of viral vectors are paramount as lentiviral vectors move into human clinical trials. This study investigates the packaging relationship between gene transfer (vector) and Gag-Pol expression constructs of HIV-1, HIV-2 and SIV. Cross-packaged vectors expressing GFP were assessed for RNA packaging, viral vector titre and their ability to transduce rat primary glial cell cultures and human neural stem cells.ResultsHIV-1 Gag-Pol demonstrated the ability to cross package both HIV-2 and SIV gene transfer vectors. However both HIV-2 and SIV Gag-Pol showed a reduced ability to package HIV-1 vector RNA with no significant gene transfer to target cells. An unexpected packaging relationship was found to exist between HIV-2 and SIV with SIV Gag-Pol able to package HIV-2 vector RNA and transduce dividing SV2T cells and CNS cell cultures with an efficiency equivalent to the homologous HIV-1 vector however HIV-2 was unable to deliver SIV based vectors.ConclusionThis new non-reciprocal cross packaging relationship between SIV and HIV-2 provides a novel way of significantly increasing bio-safety with a reduced sequence homology between the HIV-2 gene transfer vector and the SIV Gag-Pol construct thus ensuring that vector RNA packaging is unidirectional.
Highlights
Lentiviral vectors have shown immense promise as vehicles for gene delivery to non-dividing cells to cells of the central nervous system (CNS)
We investigated the cross packaging ability of the Gag-Pol components of HIV-1, HIV-2 and SIV and found a unique non-reciprocal packaging relationship between SIV Gagpol and vectors based on HIV-2
We have identified a non reciprocal cross packaging relationship between SIV Gag-Pol and a HIV-2 based GFP vector, which demonstrated equivalent transduction efficiencies in 293T cells, rat primary astrocytes and embryonic stem cells as that of homologous HIV-1 and HIV-2 vector systems
Summary
Lentiviral vectors have shown immense promise as vehicles for gene delivery to non-dividing cells to cells of the central nervous system (CNS). Improvements in the biosafety of viral vectors are paramount as lentiviral vectors move into human clinical trials. This study investigates the packaging relationship between gene transfer (vector) and Gag-Pol expression constructs of HIV-1, HIV-2 and SIV. Cross-packaged vectors expressing GFP were assessed for RNA packaging, viral vector titre and their ability to transduce rat primary glial cell cultures and human neural stem cells. Viral vectors based on primate and non-primate lentiviruses have been shown to be efficient for gene delivery to a variety of cell types both in vitro and in vivo and may offer considerable advantages in gene therapy strategies [1,2]. ∆ Env (550bp) rev tat vif vpr vpx ∆ Env (1153bp) Lentiviral vectors can provide stable gene expression following integration into the host chromosome and pseudotyping of these vectors with heterologous envelopes (page number not for citation purposes)
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