A Rapidly Maturing Far-Red Derivative of DsRed-Express2 for Whole-Cell Labeling

Fluorescent proteins (FPs) with far-red excitation and emission are desirable for multicolor labeling and live-animal imaging. We describe E2-Crimson, a far-red derivative of the tetrameric FP DsRed-Express2. Unlike other far-red FPs, E2-Crimson is noncytotoxic in bacterial and mammalian cells. E2-C...

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Bibliographic Details
Published in:Biochemistry (Easton) 2009-09, Vol.48 (35), p.8279-8281
Main Authors: Strack, Rita L, Hein, Birka, Bhattacharyya, Dibyendu, Hell, Stefan W, Keenan, Robert J, Glick, Benjamin S
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Cellular Structures
Diagnostic Imaging
Flow Cytometry
HeLa Cells
Humans
Luminescent Proteins
Microscopy, Fluorescence - methods
Microscopy, Fluorescence, Multiphoton
Protein Engineering - methods
Rapid Report
Recombinant Fusion Proteins
Red Fluorescent Protein
Spectrometry, Fluorescence - methods
Spectrometry, Fluorescence - statistics & numerical data
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Summary:Fluorescent proteins (FPs) with far-red excitation and emission are desirable for multicolor labeling and live-animal imaging. We describe E2-Crimson, a far-red derivative of the tetrameric FP DsRed-Express2. Unlike other far-red FPs, E2-Crimson is noncytotoxic in bacterial and mammalian cells. E2-Crimson is brighter than other far-red FPs and matures substantially faster than other red and far-red FPs. Approximately 40% of the E2-Crimson fluorescence signal is remarkably photostable. With an excitation maximum at 611 nm, E2-Crimson is the first FP that is efficiently excited with standard far-red lasers. We show that E2-Crimson has unique applications for flow cytometry and stimulated emission depletion (STED) microscopy.
ISSN:0006-2960
1520-4995
DOI:10.1021/bi900870u