
Abstract 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.
10120 Department of Chemistry, Tyrosine 3-Monooxygenase, Ultraviolet Rays, CHO Cells, Fungal Proteins, Cricetulus, 1311 Genetics, Genes, Reporter, Cricetinae, 540 Chemistry, Genetics, Animals, Humans, Picolinic Acids, Absorption, Radiation, Recombinant Proteins, HEK293 Cells, Spectrometry, Fluorescence, Microscopy, Fluorescence, FOS: Biological sciences, Oxygenases, Methods Online, Single-Cell Analysis
10120 Department of Chemistry, Tyrosine 3-Monooxygenase, Ultraviolet Rays, CHO Cells, Fungal Proteins, Cricetulus, 1311 Genetics, Genes, Reporter, Cricetinae, 540 Chemistry, Genetics, Animals, Humans, Picolinic Acids, Absorption, Radiation, Recombinant Proteins, HEK293 Cells, Spectrometry, Fluorescence, Microscopy, Fluorescence, FOS: Biological sciences, Oxygenases, Methods Online, Single-Cell Analysis
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