Design of HIV Vectors for Efficient Gene Delivery into Human Hematopoietic Cells

Abstract

Vectors derived from human immunodeficiency virus (HIV) hold promise for efficient gene delivery into human hematopoietic cells. In this study, HIV vectors containing different combinations of cis-acting elements, including the HIV central flap sequence, and the woodchuck posttranscriptional regulatory element (WPRE) in combination with two different promoters, were used to transduce primary human lymphocytes and cord blood CD34+ progenitor cells. The effect of these elements on the transduction efficiency and transgene expression was systematically evaluated. The results demonstrate that with the combination of flap, WPRE sequences, and the promoter derived from spleen focus-forming virus (SFFV), a foreign gene can be efficiently delivered into primary human T lymphocytes and cord blood CD34+ cells. The study establishes the parameters for proper vector design to efficiently deliver foreign genes into human hematopoietic cells.

Inhibition of HIV-1 replication with designed miRNAs expressed from RNA polymerase II promoters

“…The shRNA gene was placed under the control of the H1 promoter and inserted into pHIV7/SFgreen fluorescent protein (GFP), a third-generation HIV vector containing the GFP gene controlled by the promoter of spleen focus-forming virus (SFFV). 26 H9 cells were transduced with these vectors (HIV7/H1-tr and HIV7/H1-gag, see Figures 2b and 4a) and challenged with infectious HIV-1 NL4-3 . Although siRNAexpressing cells efficiently inhibited HIV replication at a low challenge dose, 27 HIV-1 escape mutants were observed at a higher challenge dose irrespective of the siRNA expressed (Figure 1a).…”

HIV-1 RRE RNA acts as an RNA silencing suppressor by competing with TRBP-bound siRNAs

“…16 We therefore evaluated whether a lentiviral vector derived from HIV-1, from which viral proteins are not expressed, could inhibit RNAi activity. Using the pHIV7-tdTomato vector, derived from pHIV7-CGFP harbouring both TAR and RRE, 57 we assessed RNAi activity using the same let7-luc reporter in comparison to HIV-1 NL4-3 (Fig. 5A).…”

Gene therapy to inhibit xenoantibody production using lentiviral vectors in non-human primates

“…[33][34][35][36][37] We elected to use SIVderived lentiviral vectors for this study based on reports that they are more efficient for transduction of simian stem cells. 19,20,[38][39][40] Our results using SIV-based aGT constructs are consistent with those findings.…”