High-titer purified LV-dog NIS-PGK-Puro lentivirus stocks

Promoters: Spleen focus-forming virus (SFFV) promoter for high constitutive gene expression in numerous cell types. Unlike the CMV promoter, we find that SFFV works very well in animal models. Mouse phosphoglycerate kinase I (PGK) promoter drives puromycin resistance for selection of high dNIS-expressing transduced cells.

Transgenes: Canine sodium iodide symporter (dNIS) and the puromycin resistance gene (Puro).

Envelope/Tropism: VSV.G pseudotyped HIV-1 based lentiviral vector capable of transducing a broad range of cell types and species.

Construct: This is a self-inactivating (SIN) vector in which the viral enhancer and promoter have been deleted. Thus, transcription inactivation of the LTR in the SIN provirus increases biosafety by preventing mobilization of replication competent viruses and enables regulated expression of the genes from the internal promoters without cis-acting effects of the LTR (Miyoshi et al., J Virol. 1998).

Titer: Approximately 0.5-2e7 TU/mL (by qPCR titration of transduced HeLa H1 cells)

Proposed Use: In vitro transduction of primary cells and cell lines.

Vector Map:

Iodine (¹²⁵I) Uptake Assay of LV-dNIS-PGK-Puro transduced HeLa H1 cells: