Journal article

Publication year: 2022

Journal: Scientific Reports

Volume number: 12

Issue number: 1

ISSN: 2045-2322

Open access status: Gold

DOI Link: https://doi.org/10.1038/s41598-022-09169-x

Publisher: Nature Research

Abstract: 
Modern biology investigations on phytochromes as near-infrared fluorescent pigments pave the way for the development of new biosensors, as well as for optogenetics and in vivo imaging tools. Recently, near-infrared fluorescent proteins (NIR-FPs) engineered from biliverdin-binding bacteriophytochromes and cyanobacteriochromes, and from phycocyanobilin-binding cyanobacterial phytochromes have become promising probes for fluorescence microscopy and in vivo imaging. However, current NIR-FPs typically suffer from low fluorescence quantum yields and short fluorescence lifetimes. Here, we applied the rational approach of combining mutations known to enhance fluorescence in the cyanobacterial phytochrome Cph1 to derive a series of highly fluorescent variants with fluorescence quantum yield exceeding 15%. These variants were characterised by biochemical and spectroscopic methods, including time-resolved fluorescence spectroscopy. We show that these new NIR-FPs exhibit high fluorescence quantum yields and long fluorescence lifetimes, contributing to their bright fluorescence, and provide fluorescence lifetime imaging measurements in E.coli cells.

Harvard Citation style: Nagano, S., Sadeghi, M., Balke, J., Fleck, M., Heckmann, N., Psakis, G., et al. (2022) Improved fluorescent phytochromes for in situ imaging, Scientific Reports, 12(1), Article 5587. https://doi.org/10.1038/s41598-022-09169-x

APA Citation style: Nagano, S., Sadeghi, M., Balke, J., Fleck, M., Heckmann, N., Psakis, G., & Alexiev, U. (2022). Improved fluorescent phytochromes for in situ imaging. Scientific Reports. 12(1), Article 5587. https://doi.org/10.1038/s41598-022-09169-x