A noncytotoxic DsRed variant for whole-cell labeling

Rita L. Strack; Daniel E. Strongin; Dibyendu Bhattacharyya; Wen Tao; Allison Berman; Hal E. Broxmeyer; Robert J. Keenan M.D.; Benjamin S. Glick

doi:10.1117/12.808046

16 February 2009 A noncytotoxic DsRed variant for whole-cell labeling

Rita L. Strack, Daniel E. Strongin, Dibyendu Bhattacharyya, Wen Tao, Allison Berman, Hal E. Broxmeyer, Robert J. Keenan M.D., Benjamin S. Glick

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Proceedings Volume 7191, Fluorescence In Vivo Imaging Based on Genetically Engineered Probes: From Living Cells to Whole Body Imaging IV; 719109 (2009) https://doi.org/10.1117/12.808046
Event: SPIE BiOS, 2009, San Jose, California, United States

Abstract

Fluorescent proteins (FPs) are extremely useful tools for whole-cell, tissue, and animal labeling. For these purposes, FPs may be monomeric or oligomeric, but should meet the criteria of being tolerated at high expression levels in cells and having desirable photophysical properties. Our goal was to create a variant of DsRed-Express that maintains the brightness, fast-maturation, and photostability of this protein, while exhibiting decreased cytotoxicity. For this purpose, we mutated the surface of DsRed-Express to decrease aggregation and created DsRed-Express2. DsRed-Express2 retains the favorable photophysical properties of DsRed-Express while showing dramatically reduced cytotoxicity and higher expression in bacterial and mammalian systems. Further, it was shown that DsRed-Express2 outperforms other red FPs as a label for bacterial and mammalian cells.

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Rita L. Strack, Daniel E. Strongin, Dibyendu Bhattacharyya, Wen Tao, Allison Berman, Hal E. Broxmeyer, Robert J. Keenan M.D., and Benjamin S. Glick "A noncytotoxic DsRed variant for whole-cell labeling", Proc. SPIE 7191, Fluorescence In Vivo Imaging Based on Genetically Engineered Probes: From Living Cells to Whole Body Imaging IV, 719109 (16 February 2009); https://doi.org/10.1117/12.808046

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KEYWORDS

Luminescence

Flow cytometry

Proteins

Signal detection

Bone

Computer programming

Crystals

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