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A noncytotoxic DsRed variant for whole-cell labeling
Rita L Strack1, Daniel E Strongin, Dibyendu Bhattacharyya
1Department of Biochemistry and Molecular Biology, The University of Chicago, Gordon Center W238, Chicago, Illinois 60637, USA.
Nature Methods
|October 28, 2008
Summary
Researchers developed DsRed-Express2, a novel red fluorescent protein (RFP). This engineered protein shows minimal cytotoxicity and superior photostability, making it ideal for cell labeling applications.
Area of Science:
- Biochemistry
- Cell Biology
- Molecular Biology
Background:
- Fluorescent proteins are widely used for cell labeling.
- Many red fluorescent proteins (RFPs) cause cytotoxicity with high-level expression systems.
Purpose of the Study:
- To engineer a novel, rapidly maturing tetrameric red fluorescent protein with minimal cytotoxicity.
- To evaluate the performance of the engineered RFP in bacterial and mammalian cells.
Main Methods:
- Engineering of a tetrameric fluorescent protein (DsRed-Express2).
- Assessment of cytotoxicity in bacterial and mammalian cell expression systems.
- Evaluation of expression stability, photostability, and phototoxicity compared to other RFPs.
Main Results:
- DsRed-Express2 demonstrated minimal cytotoxicity.
- The engineered protein showed strong and stable expression in both bacterial and mammalian cells.
- DsRed-Express2 outperformed existing RFPs in photostability and phototoxicity.
Conclusions:
- DsRed-Express2 is a viable alternative to existing RFPs for cell labeling.
- The engineered protein offers improved performance regarding cytotoxicity and photostability.
- This advancement facilitates robust cell labeling in various biological systems.
