U2OS Cells GFP-NUP98,

Application

U2OS Cells have been used to assess cell proliferation using Alamar Blue and live cell imaging. This product is a human U2OS cell line in which the genomic NUP98 gene has been endogenously tagged with a 3XFLAG^™ epitope and a Green Fluorescent Protein (GFP) gene using CompoZr^® Zinc Finger Nuclease technology. Integration resulted in endogenous expression of a fusion protein in which the 3XFLAG is attached to the N-terminus of GFP and the GFP is attached to the N-terminus of NUP98. Fluorescence imaging shows characteristic NUP98 nuclear 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

Features and Benefits

Zinc finger nuclease (ZFN)-mediated targeted integration of a fluorescent GFP tag just behind the start codon of the NUP98 gene on chromosome 11p15.4 to create a cell line exhibiting stable expression of the transgene tagged with GFP on the N-terminus of the protein.The U2OS cells are adherent, with a doubling time of approx. 29 hours.

General description

U2OS GFP-NUP98 are bone osteosarcoma cells from a human 15 year old caucasian female having a ZFN modification creating a GFP-NUP98 transgene expressed from the endogenous NUP98 gene locus.This cell line was derived from ATCC Catalog No. HTB-96.

Legal Information

3xFLAG is a trademark of Sigma-Aldrich Co. LLC

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 5A 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 (v/v) of 10%.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.