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

 

Promoters: Spleen focus-forming virus (SFFV) for high constitutive NIS gene expression and phosphoglycerate kinase (PGK) 

Transgenes: Rhesus sodium iodide symporter (RhNIS) driven by the SFFV promoter and puromycin (Puro) antibiotic resistance gene driven by the PGK promoter

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 ready-to-use lentivirus preparation. The virus encodes the rhesus sodium iodide symporter (RhNIS) cDNA under control of the spleen focus-forming virus (SFFV) promoter and the puromycin resistance gene (Puro) under the phosphoglycerate kinase (PGK) promoter (see below). The lentiviral vectors are self-inactivating (SIN) vectors in which the viral enhancer and promoter have been deleted. Transcription inactivation of the LTR in the SIN provirus increases biosafety by preventing mobilization by 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:

NIS Expression: HeLaH1 cells were transduced with LV-RhNIS-PGK-Puro (MOI = 40). Uptake of ¹²⁵I by 2 x 10⁵ transduced cells was assayed in the presence or absence of KClO₄, an inhibitor of NIS-mediated ¹²⁵I uptake.