Figure 1

Lentiviral transduction of immortalized sensory neuronal cells (50B11) in vitro. A. Schematic representation of the lentivectors. EGFP-expressing lentivector (LV) plasmids carrying a panel of various recombinant promoters were generated. Cis-acting sequences included were cPPT (central polypurine tract) to enhance transduction efficiency, and WPRE (woodchuck posttranscriptional regulatory element) to improve transgene expression. B, C and D. LV transduction efficiencies in 50B11 cells (MOI = 2) were assessed by FACS analysis. B. Transduction efficiencies were compared between vectors with different constitutively active cellular promoters (n = 4 experiments/promoter), including: 1) human elongation factor 1α (EF1α); 2) hybrid promoter containing a human CMV enhancer element 5' to the chicken β-actin promoter (CAG); 3) human ubiquitin C (UbC); and 4) murine phosphoglycerate kinase gene (PGK). C. Transduction efficiencies using LVs with (first-generation, LV1-EF1α-EGFP) or without viral accessory proteins (second-generation, LV2-EF1α-EGFP) were compared (n = 8 experiments/packaging system). D. Alternative pseudotyping of LVs was performed to determine its effects on transduction efficiency (n = 4 experiments/pseudotype). The envelope coat proteins analyzed were as follows: 1) vesicular stomatitis virus G protein (VSV-G); 2) rabies SAD glycoproteins (RABSAD); 3) glycoprotein from rabies virus PV strain (RABPV); and 4) lymphochoriomeningitis virus envelope (LCMV). Post hoc differences between vectors are represented by bars; * p < 0.001.