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Genetically encoded betaxanthin-based small-molecular fluorescent reporter for mammalian cells.

  • Stücheli P Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, CH-4058 Basel, Switzerland.
  • Sieber S Department of Chemistry, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland.
  • Fuchs DW Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, CH-4058 Basel, Switzerland.
  • Scheller L Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, CH-4058 Basel, Switzerland.
  • Strittmatter T Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, CH-4058 Basel, Switzerland.
  • Saxena P Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, CH-4058 Basel, Switzerland.
  • Gademann K Department of Chemistry, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland.
  • Fussenegger M Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, CH-4058 Basel, Switzerland.
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  • 2020-05-19
Published in:
  • Nucleic acids research. - 2020
Absorption, Radiation
Animals
CHO Cells
Cricetinae
Cricetulus
Fungal Proteins
Genes, Reporter
HEK293 Cells
Humans
Microscopy, Fluorescence
Oxygenases
Picolinic Acids
Recombinant Proteins
Single-Cell Analysis
Spectrometry, Fluorescence
Tyrosine 3-Monooxygenase
Ultraviolet Rays
English We designed and engineered a dye production cassette encoding a heterologous pathway, including human tyrosine hydroxylase and Amanita muscaria 4,5-DOPA dioxygenase, for the biosynthesis of the betaxanthin family of plant and fungal pigments in mammalian cells. The system does not impair cell viability, and can be used as a non-protein reporter system to directly visualize the dynamics of gene expression by profiling absorbance or fluorescence in the supernatant of cell cultures, as well as for fluorescence labeling of individual cells. Pigment profiling can also be multiplexed with reporter proteins such as mCherry or the human model glycoprotein SEAP (secreted alkaline phosphatase). Furthermore, absorbance measurement with a smartphone camera using standard application software enables inexpensive, low-tech reporter quantification.
Language
  • English
Open access status
gold
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Persistent URL
https://sonar.ch/global/documents/144520
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