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VV(Li)-RucGFP-hNIS-tRFP

  • 0.25 mL / Standard (OV4001) $ 750

Ready to use vaccinia virus expressing Ruc-GFP, hNIS, and tRFP.

  • Description: Replication competent vaccinia virus encoding Renilla luciferase (Ruc)-green fluorescent protein (GFP) fusion, human sodium iodide symporter (hNIS), and TurboFP635-like protein (tRFP). These additional transcription units are inserted into, and disrupt the function of, the viral F14.5L, J2R (thymidine kinase) and A56R (hemagglutinin) genes, respectively (see diagram below).
  • Strain: Lister strain
  • Transgenes: Green fluorescent protein (GFP), human sodium iodide symporter (hNIS), TurboFP635-like protein (tRFP), and Renilla luciferase (Ruc).
  • Biosafety Level: 2
  • Titer: Standard (clarified supernatant, at least 5e6 TCID50/ml).
  • Propagation method: Inoculate producer cells (e.g. Vero) at an MOI of 0.1 for 2-3 hours. After 48 hours harvest culture supernatant and cells and freeze-thaw or sonicate 3X. Clarify the supernatant by low-speed centrifugation and purify through a  sucrose cushion.  Resuspend the viral pellet in 1mM TrisCl, pH 9.
  • Proposed Use: Ready to use viral stock for propagation and cell killing assays.
  • USDA permit required for shipping? Yes. Click here to fill out Application Form VS 16-3 to obtain a USDA APHIS VS 16-6 or 16-6A permit.

Transgene Validation

Fluorescence: Vero cells were mock-infected (A-C) or infected with VV(Li)RucGFP-hNIS-tRFP MOI 0.1 (D-F) and imaged at 48 h.p.i.

NIS Function: Vero cells were mock-infected or infected with VV(Li)-RucGFP-hNIS-tRFP at the indicated MOI. After 24 hours, uptake of 125I was measured in the presence or absence of KClO4, an inhibitor of NIS-mediated 125I uptake.

Luciferase Assay: Vero cells were mock-infected or infected with VV(Li)-RucGFP-hNIS-tRFP at an MOI of 0.1 or 1. After 48 hours, 1.25 µg of coelenterazine was added to each of the wells and luminescence (RLU) was immediately measured using a microplate reader.

 

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      Use Fewer Animals

      Our Reduction Campaign

      At Imanis, we are committed to promoting the practice of the 3Rs in animal research. Learn how we are decreasing the use of animals and research as well as saving up to 15% on your orders. Continue reading...