Species: Human

Cell type: Colorectal carcinoma

Transgenes: Near-infrared fluorescent protein (iRFP; ex/em = 690/713) with puromycin resistance (Puro) for selection with puromycin.

Media: DMEM, 10% FBS, 1% Pen/Strep, 1 μg/mL puromycin

Description: HCT116-iRFP-Puro is a polyclonal population of the human colorectal carcinoma cell line HCT116 (ATCC® CCL-247™) transduced with a lentiviral vector LV-iRFP-P2A-Puro (LV032) encoding the near-infrared fluorescent protein (iRFP) cDNA under the SFFV promoter and the puromycin resistance gene (Puro) via a P2A cleavage peptide.

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 HCT116-iRFP-Puro cell line has been tested for mycoplasma contamination and is certified mycoplasma free.

Cell Line Authentication: HCT116 cell line was authenticated are certified free of interspecies cross-contamination by short tandem repeat (STR) profiling with 9 STR loci including CSF1PO, D13S317, D16S539, D5S818, D7S820, TH01, TPOX, vWA and sex chromosome marker Amelogenin.

Recommended uses: In vitro: This is a high iRFP expressing cell line suitable for use as a positive control cell line in fluorescence assays to verify iRFP expression in your lentiviral transduced cells. In vivo: This human colorectal cell line grows as xenografts in immunocompromised mice. Growth of these tumors can be monitored in vivo using noninvasive optical imaging for iRFP fluorescence.

Note: Please ensure that your optical imaging system has the appropriate filter sets for imaging iRFP (iRFP; ex/em = 690/713)

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

Flow Cytometry for iRFP: HCT116-iRFP-Puro (red) or control (HCT116; grey) cells were fixed with paraformaldehyde and analyzed by flow cytometry (20,000 events).