Exp Mol Med.  2001 Sep;33(3):174-178.

Monitoring transfected cells without selection agents by using the dual-cassette expression EGFP vectors

• Kang SG
• Lee EK
• Schaus S
• Henderson

Abstract

Conventional methods of selecting gene transfected cells by toxic agents may yield ambiguous results. It is difficult to determine whether cell death is due to selection agents or gene transfection, owing to the substantial overlap of the time-courses for both effects. Therefore, to determine transfection-induced cell toxicity, the mammalian expression vector pEGFP-N1 (CLONTECH Lab., Palo Alto, CA, USA) has been modified to the dual-cassette expression vectors named pEGFP-Ks by the relocation of its EGFP expression cassette. We have precisely monitored the cells transfected with this vector on our custom culture dishes, thereby bypassing the need for selection agent or fluorescent cell sorting. This is a useful method to screen genes encoding potential toxic or useful proteins without performing undesirable selection agent and also can be used to monitor the transfected cells for various purposes, either the inhibition or proliferation of mammalian cells for applications in biotechnology.

Keyword

fluorescent protein; EGFP; cancer cell proliferation; transfection; dual-cassette expression vector

MeSH Terms

Cell Culture/methods
Cell Death
Genes, Reporter
*Genetic Vectors
Human
Indicators and Reagents
Luminescent Proteins/*genetics/metabolism
Recombinant Fusion Proteins/genetics/metabolism
Telomerase/genetics/metabolism
Transfection/*methods