Genetically encoded betaxanthin-based small-molecular fluorescent reporter for mammalian cells. Issue 12 (18th May 2020)

• Record Type: Journal Article
• Authors: Stücheli, Pascal; Sieber, Simon; Fuchs, David W; Scheller, Leo; Strittmatter, Tobias; Saxena, Pratik; Gademann, Karl; Fussenegger, Martin
• Is Part Of: Nucleic acids research. Volume 48:Issue 12(2020)
• Page Start: e67
• Page End: e67
• Publication Date: 2020-05-18
• Subjects: Nucleic acids -- Periodicals; Molecular biology -- Periodicals; 572.805
• DOI: 10.1093/nar/gkaa342
• Languages: English
• ISSNs: 0305-1048

Abstract

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.