Species: Mouse

Cell type: Mammary carcinoma

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

Media: RPMI, 10% FBS, 1% Pen/Strep, 2 ug/mL puromycin

Description:

4T1-eGFP-Puro is a polyclonal population of the murine mammary carcinoma cell line 4T1 (ATCC® CRL-2539™) 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 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:

The parental 4T1 cell line was authenticated and cells are certified free of interspecies cross-contamination by short tandem repeat (STR) profiling with 27 STR loci. 

Recommended uses:

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

In vivo: 4T1 cells form tumors post implantation into immunocompromised or syngenic Balb/c mice. These tumors can be detected using optical imaging for GFP fluorescence at post mortem analysis.

Note: In-life imaging for GFP fluorescence is not recommended due to high background autofluorescence and low light penetration. Tissues may be harvested post mortem for analysis by conventional microscopy. For in vivo imaging, please use luciferase or the nonimmunogenic murine NIS reporter gene.

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

Transgene Validation:

Flow cytometry of 4T1-eGFP-Puro (green) and an isotype control (gray)

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.