SKOV3 Cells GFP-HER2

General description

SKOV3 GFP-HER2 are ovarian adenocarcinoma cells from a human 64 year old caucasion female having a ZFN modification creating a HER2-GFP transgene expressed from the endogenous HER2 gene locus.

This cell line was derived from ATCC Catalog No. HTB-77.

Application

SKOV3 Cells GFP-HER2 has been used for live-cell screening assays to find compounds that affect cell structure or signaling pathways.

This product is a human SKOV3 cell line in which the genomic HER2 gene has been endogenously tagged with a Green Fluorescent Protein (GFP) gene using CompoZr^® Zinc Finger Nuclease technology. Integration resulted in endogenous expression of the fusion protein in which GFP is attached to the C-terminus of HER2. Fluorescence imaging shows characteristic HER2 membrane expression. This stable cell line was expanded from a single clone. The target′s gene regulation and corresponding protein function are preserved in contrast to cell lines with overexpression via an exogenous promoter.

To learn more, please visit the Cellular Reporter Cell Line webpage

Biochem/physiol Actions

Human epidermal growth factor receptor 2 (HER2) signaling pathway promotes cell proliferation and survival in majority of breast cancers. Thus, overexpression of this protein leads to breast cancer. Heterodimeric complex of HER2 and phosphatidylinositide 3-kinase (PI3K) is the most potent stimulator of the phosphatidylinositol-3-kinase (PI3K)/Akt anti-apoptosis pathway. Upregulated expression of HER2 is associated with the development of ovarian, colorectal, pancreatic, endometrial and gastric cancers. Trastuzumab, an antibody, works by binding to a domain in the external domain of HER2. This domain is missing in p95, a truncated form of HER2, and hence these cancer cells show resistance to trastuzumab. HER2 protein can be used as a prognostic marker and as a therapeutic option for gynecologic cancers.

Features and Benefits

Zinc Finger Nuclease (ZFN)-mediated targeted integration of a fluorescent GFP tag into the last exon of the HER2 gene on chromosome 17q21.1 to create a cell line exhibiting stable expression of the transgene tagged with GFP on the C-terminus of the protein.

The SKOV3 cells are adherent, with a doubling time of approx. 48 hours.

Quality

Tested for Mycoplasma, sterility, post-freeze viability, short terminal repeat (STR) analysis for cell line identification, cytochrome oxidase I (COI) analysis for cell line species confirmation.

Preparation Note

Media Renewal changes two to three times per week.

Rapidly thaw vial by gentle agitation in 37°C water bath (~2 minutes), keeping vial cap out of the water. Decontaminate with 70% ethanol, add 9 mL culture media and centrifuge 125 x g (5-7 minutes). Resuspend in complete culture media and incubate at 37°C in a 5% CO2 atmosphere.

Subculture Ratio: approx. 1:3-1:6

The base medium for this cell line is McCoy′s 5A Medium, Cat. No. M8403. To make the complete growth medium, add the following components to the base medium: fetal bovine serum, Cat. No. F2442, to a final concentration (v/v) of 10% and L-glutamine, Cat. No. G7513, at a final concentration of 1.5 mM.

Cell freezing medium-DMSO 1X, Cat. No. C6164.

Legal Information

CompoZr is a registered trademark of Sigma-Aldrich Co. LLC