Species: Mouse

Cell type: Colorectal Cancer

Transgenes: Enhanced green fluorescent protein (eGFP) with puromycin resistance (Puro) for selection with puromycin

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

Description: CT26.WT-eGFP-Puro is a polyclonal population of the murine colorectal carcinoma cell line CT26.WT (ATCC® CRL-2638™) transduced with LV-eGFP-PGK-Puro (LV031) encoding enhanced green fluorescent protein (eGFP) cDNA under the spleen focus-forming virus (SFFV) promoter and the puromycin resistance gene (Puro) under control of the 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).

Cell Line Authentication: Authentication of the parental CT26.WT cell line was performed by short tandem repeat (STR) profiling with 27 STR loci. STR profiling of CT26.WT cells are verified and there is no interspecies cross contamination detected.

Recommended uses:

In vitro: This is a high eGFP expressing cell line suitable for use as a positive control cell line in fluorescence assays to verify GFP expression in your lentiviral transduced cells.

In vivo: CT26.WT cells form tumors post implantation into immunocompromised or Balb/c syngeneic mice. 

Note: In-life imaging for GFP fluorescence is not recommended due to high background autofluorescence. Tissues may be harvested post mortem for analysis by conventional microscopy. For in vivo imaging in immunocompetent animals, please consider using the nonimmunogenic murine NIS reporter gene.

Publications that used associated reagents: 

LV-eGFP-PGK-Puro (LV031): Shen et al. Immunovirotherapy with vesicular stomatitis virus and PD-L1 blockade enhances therapeutic outcome in murine acute myeloid leukemia. Blood. 2016. March 17: 127(11): 1449-58.

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

Transgene Validation:

Flow cytometry of CT26.WT-GFP-Puro (green) and control CT26.WT (grey) cells.