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CT26.WT-iRFP-Neo/mNIS-Puro

Species: Mouse

Cell type: Colorectal Cancer

Transgenes: Near infrared fluorescent protein (iRFP; ex/em = 690/713 nm) with neomycin resistance (Neo) for selection with G418 and murine sodium iodide symporter (mNIS) with puromycin resistance (Puro) for selection with puromycin.

Media: DMEM, 10% FBS, 1% Pen/Strep, 0.4 mg/mL G418, 3 μg/mL puromycin

Description:

CT26.WT-iRFP-Neo/mNIS-Puro is a polyclonal population of the colorectal carcinoma cell line CT26.WT (ATCC® CRL-2638™) transduced with 1) LV-iRFP-P2A-Neo (LV033) encoding the near-infrared fluorescent protein (iRFP) cDNA under the spleen focus-forming virus (SFFV) promoter linked to the neomycin resistance gene (Neo) via a P2A cleavage peptide and 2) LV-mNIS-PGK-Puro (LV022) encoding the murine sodium iodide symporter (mNIS) cDNA under the spleen focus-forming virus (SFFV) promoter and the puromycin resistance gene (Puro) under the mouse phosphoglycerate kinase (PGK) promoter.

The lentiviral vector used is a self-inactivating (SIN) vector in which the viral enhancer and promoter has 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).

Mycoplasma Testing:

The CT26.WT-iRFP-Neo/mNIS-Puro cell line has been tested for mycoplasma contamination and is certified mycoplasma free.

Cell Line Authentication:

The parental CT26.WT cell line was authenticated and certified free of interspecies cross-contamination by short tandem repeat (STR) profiling with 27 STR loci.

Recommended uses:

In vitro: This is a high iRFP and NIS expressing cell line suitable for use as a positive control cell line in fluorescence and iodine uptake assays to verify iRFP or NIS expression in your lentiviral transduced cells.

In vivo: CT26.WT cells form tumors post implantation in mice. The in vivo growth of these tumors can be monitored using noninvasive optical imaging for iRFP or noninvasive, high-resolution 3D PET or SPECT imaging for NIS.

Morphology: Low- and high-density cell morphology (200x)

NIS Function Assay (Iodine Uptake): Cells were incubated with I-125 for 1h in the presence or absence of KClO4, an inhibitor of NIS mediated iodine uptake. Radioiodine concentrated within the cells was measured with a gamma counter.

Flow Cytometry for iRFPCT26.WT-iRFP-Neo/mNIS-Puro cells (red) or control (CT26.WT-Fluc-Neo; grey) cells were fixed with paraformaldehyde and analyzed by flow cytometry (20,000 events).

Why Choose Imanis?

With quality reagents, services, and support, we have you covered at all stages of your study. Our expert scientists can help you select the best reagents for your needs and offer tips to optimize your experiments and achieve superior results. Our wide-range of reporter gene and oncolytic virus reagents undergo rigorous quality control testing so you can be confident in the quality of the products you receive. And with our competitive prices, you can complete your study for less.

  • “Our group has, and continues to, use NIS as a noninvasive reporter for cell transplantation studies in mice and in pigs. Imanis has provided expert technical and analytical support for this research, and has allowed us to publish our research in high impact journals, including Science Translational Medicine.” – Dr. Raymond Hickey, Mayo Clinic

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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...