A549 Cells RFP-STAT3,

Application

This product is a human A549 cell line in which the genomic STAT3 gene has been endogenously tagged with a Red Fluorescent Protein (RFP) gene using CompoZr^® zinc finger nuclease technology. Integration resulted in endogenous expression of the fusion protein in which RFP is attached to the N-terminus of STAT3. Fluorescence imaging shows RFP-STAT3 expression throughout the cell. Upon activation with the ligand IL-6, the cell line shows redistribution of STAT3 to the nucleus, making it useful for high content screening of compounds that modulate STAT3 activity. For example, the redistribution can be abolished by a selective inhibitor of STAT3, Stattic. 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

Features and Benefits

Zinc Finger Nuclease (ZFN)-mediated targeted integration a fluorescent RFP tag just behind the start codon of the STAT3 gene on chromosome 17q21.2 to create a cell line exhibiting stable expression of the transgene tagged with GFP on the N-terminus of the protein.These A549 cells are adherent with a doubling time of approx. 22 hours.

General description

A549 RFP-STAT3 are lung carcinoma, epithelial cells from a human 58 year old male caucasion having a ZFN modification creating an RFP-STAT3 transgene expressed from the endogenous STAT3 gene locus.This cell line was derived from ATCC catalog No. CCL-185.

Legal Information

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

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. 1:3-1:6The base medium for this cell line is RPMI-1640, Cat. No. R0883. 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 2 mM.Cell freezing medium-DMSO 1X, Cat. No. C6164.

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.