U2OS GFP-ACTB,

Application

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

This product is a human U2OS cell line in which the genomic ACTB 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 N-terminus of actin. Fluorescence imaging shows that the actin can polymerize to form the characteristic pattern of actin filaments. 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.

Biochem/physiol Actions

Actin β (ACTB) is an abundant cytoskeletal housekeeping protein. The protein, expressed in the nucleus, controls gene expression, cell division and proliferation. ACTB is generally used as a reference gene in measuring expression levels in tumors.

β-Actin modulates cell growth, migration and the G-actin pool. Mutations in ACTB leads to pleiotropic developmental disorder. GFP (green fluorescent protein) is used for quantitative analysis.

Components

U2OS GFP-ACTB cells

Features and Benefits

The U2OS cells are adherent, with a doubling time of approx. 29 hours.

GFP-tagged on chromosome 7p22.1

General description

U2OS GFP-ACTB are osteosarcoma, epithelial cells, from a human caucasian female (aged 15 years), with a GFP-tagged modification.

This product was derived from ATCC Cat. No. HTB-96.ACTB (actin beta) gene codes for β-actin. It is a cytoskeletal housekeeping protein. It is present in the cytoplasm. The ACTB gene is located on human chromosome 7p22.1. GFP (green fluorescent protein) is a major reporter protein that has fluorescence properties. It can fluoresce without the help of any cofactors or substrates except for oxygen.

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 McCoy′s 5A1 Medium Modified, Cat. No. M9309. To make the complete growth medium, add the following components to the base medium: fetal bovine serum, Cat. No. F4135, to a final concentration of 10%.The cell freezing medium is DMSO 1X (Cat. No. C6164).

Quality

Tested for Mycoplasma, sterility, post-freeze viability, short terminal repeat (STR) analysis for cell line identification, PCR assay for cell line species confirmation.