A noncytotoxic DsRed variant for whole-cell labeling

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

Research output: Chapter in Book/Report/Conference proceedingConference contribution

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 outper forms other red FPs as a label for bacterial and mammalian cells.

Original languageEnglish (US)
Title of host publicationFluorescence In Vivo Imaging Based on Genetically Engineered Probes
Subtitle of host publicationFrom Living Cells to Whole Body Imaging IV
DOIs
StatePublished - Jun 15 2009
EventFluorescence In Vivo Imaging Based on Genetically Engineered Probes: From Living Cells to Whole Body Imaging IV - San Jose, CA, United States
Duration: Jan 25 2009Jan 26 2009

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume7191
ISSN (Print)1605-7422

Other

OtherFluorescence In Vivo Imaging Based on Genetically Engineered Probes: From Living Cells to Whole Body Imaging IV
CountryUnited States
CitySan Jose, CA
Period1/25/091/26/09

Keywords

  • Directed evolution
  • DsRed
  • Fluorescent protein
  • Whole-cell labeling

ASJC Scopus subject areas

  • Atomic and Molecular Physics, and Optics
  • Electronic, Optical and Magnetic Materials
  • Biomaterials
  • Radiology Nuclear Medicine and imaging

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  • Cite this

    Strack, R. L., Keenan, R. J., Strongin, D. E., Bhattachryya, D., Berman, A., Glick, B. S., Wen, T., & Broxmeyer, H. E. (2009). A noncytotoxic DsRed variant for whole-cell labeling. In Fluorescence In Vivo Imaging Based on Genetically Engineered Probes: From Living Cells to Whole Body Imaging IV [719109] (Progress in Biomedical Optics and Imaging - Proceedings of SPIE; Vol. 7191). https://doi.org/10.1117/12.808046