LV-eGFP-P2A-hNIS purified high titer lentivirus stock

• Description: LV-SFFV-eGFP-P2A-hNIS. 

• This is a ready-to-use lentivirus preparation. The virus encodes the enhanced green fluorescent protein (eGFP) cDNA under control of the spleen focus-forming virus (SFFV) promoter linked to the human sodium iodide symporter (hNIS) cDNA via a P2A cleavage peptide (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. 72:8150-8157).

• Titer: 1e8 TU/ml in DMEM
• Size: 0.25 mL or 1.0 mL
• Promoter: SFFV spleen focus forming virus for high constitutive gene expression in numerous cell types. Unlike the CMV promoter, SFFV works very well in human and rodent cells.
• Transgene: Enhanced GFP linked to human NIS by a P2A sequence.

Transgene Validation

Fluorescence Expression: HeLaH1 cells were transduced with LV-eGFP-P2A-hNIS at the indicated MOIs. After 6 days fluorescence expression of the transduced (green) and untransducted parental HeLaH1 cells (grey) was analyzed by flow cytometry.

 

NIS Gene Expression: HeLaH1 cells were transduced with LV-eGFP-P2A-hNIS at the indicated MOIs. After 1 week ¹²⁵I uptake by the cells was measured in the presence or absence of KClO₄, an inhibitor of NIS-mediated ¹²⁵I uptake.