Figure 1.

Stable expression of cocal and VSV-G envelopes in 293T-cells. (a) Schematic representation of cocal and vesicular stomatitis virus envelope glycoprotein (VSV-G) envelope plasmids and lentiviral transfer plasmids used for the generation of Lentiviral vector (LV) producer cells. (b) Unconcentrated LV titer from standard vector production using transient transfection (left), or from cells stably expressing each envelope (right). Mean titer is given as infectious units per ml (IU/ml) from cells transduced in triplicates. (c) SYBR Green-based quantitative PCR measurements of envelope transcripts in two independent cocal or VSV-G cell lines. Envelope mRNA expression was calculated relative to expression of the endogenous gene glyceraldehyde 3-phosphate dehydrogenase from reactions run in triplicates. Cell line A was used to generate the titers given in b. Error bar shows standard error (SE) of the mean. CMV, human cytomegalovirus promoter; hBGint, human beta globin intronic sequence; 2A,self-cleaving peptide sequence; hBpA, human beta globin polyA sequence; ΔLTR, long terminal repeat containing short deletion; RSV, Rous sarcoma virus promoter; PGK, phosphoglycerate kinase 1 promoter (human); eGFP, enhanced green fluorescent protein; Wpre, woodchuck hepatitis virus regulatory element; MSCV, murine stem cell virus promoter; TCRα, T-cell receptor alpha chain; TCRβ, T-cell receptor beta chain.