High-titer purified LV-PGK-hNIS-P2A-Puro lentivirus stocks

Promoter: Phosphoglycerate kinase (PGK) promoter for constitutive gene expression in numerous cell types. 

Reporter gene: Human sodium iodide symporter (hNIS)

Selection gene: Puromycin (Puro)

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

Description: This is a ready-to-use lentivirus preparation. The virus encodes the human sodium iodide symporter (hNIS) cDNA under control of the phosphoglycerate kinase (PGK) promoter linked to the puromycin resistance gene (Puro) via a P2A cleavage peptide (see below). 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 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 (I-125) Uptake Assay of transduced HeLa H1 cells: HeLaH1 cells were transduced with LV-PGK-hNIS-P2A-Puro (MOI = 5) and after 3 days the cells were amplified under puromycin selection. Uptake of ¹²⁵I by 3 x 10⁵ transduced cells was assayed in the presence or absence of KClO₄, an inhibitor of NIS-mediated ¹²⁵I uptake.