pLV-SFFV-Luc2-P2A-humanNIS

• Name of plasmid: pLV-SFFV-Luc2-P2A-human NIS
• Description: pHR' SIN-SFFV-Luc2-P2A-hNIS plasmid DNA
• Amount of DNA: 20 ug
• Promoter: SFFV spleen focus forming virus promoter for high constitutive gene expression in numerous cell types. Unlike the CMV promoter, SFFV works very well in human, rodent, and other animal cells.
• Transgenes: Human sodium iodide symporter (hNIS) and firefly luciferase (Luc2)
• Linker: Self-cleaving peptide P2A
• Proposed Use: For generation of lentiviral vector stocks encoding firefly luciferase and human NIS in a triple transfection protocol.

Plasmid Map:

Transgene Validation:

Luciferase Assay: 10⁴, 10⁵, or 10⁶ LV-Luc2-hNIS transduced cells were placed in wells of a 96-well plate and 0.3 mg of d-luciferin was added to the indicated wells. The plate was immediately imaged using a Xenogen IVIS Spectrum.

Uptake Assay: NIS mediated ¹²⁵I uptake of LV-Luc2-hNIS transduced cells was assayed in the presence or absence of KClO₄, a competitive inhibitor of iodine uptake.