A small and highly sensitive red/far-red optogenetic switch for applications in mammals

Zhou, Yang; Kong, Dan; Wang, Xinyi; Yu, Guiling; Wu, Xin; Guan, Ningzi; Weber, Wilfried; Ye, Haifeng

doi:10.1038/s41587-021-01036-w

Cited by 80 publications

(74 citation statements)

References 50 publications

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“…For future expression in mammalian cells it must be noted that unlike bacteriophytochromes, Cph1-based fluorophores cannot assemble with the biliverdin present in mammalian cells. However, the PCB cofactor can be fed to the cells 53 or generated from the biliverdin by co-expressing pcyA (phycocyano-bilin:ferredoxin oxidoreductase) from the transfected plasmid 54 , 55 . Expressing HY2 (phytochromobilin:ferredoxin oxidoreductase) might be possible too 56 .…”

Section: Resultsmentioning

confidence: 99%

Improved fluorescent phytochromes for in situ imaging

Nagano

Sadeghi

Balke

et al. 2022

Sci Rep

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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.

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Section: Resultsmentioning

confidence: 99%

Improved fluorescent phytochromes for in situ imaging

Nagano

Sadeghi

Balke

et al. 2022

Sci Rep

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“…The transcription of FHY1 and FHL is further controlled by transcription factors such as far-red elongated Hypocotyl 3 and far-red impaired response 1 ( 173 , 174 ). The PhyA-FHL pair was fused to GAL4 and GAL1, respectively, to develop a red light-inducible gene expression system in yeast and mammals ( 175 , 176 ).…”

Section: Overview Of Photosensory Modulesmentioning

confidence: 99%

“…Through a frequency-response analysis using different light pulses (minutes to hours), paracrine STAT3 activation was suggested to be critical for maintaining sustained Ras activation ( 234 ). Additionally, similar Opto-SOS systems using iLID-CAAX and SspB-SOScat linked by a P2A cleavable peptide ( 236 ) or using a REDMAP (red/far-red light-mediated and miniaturized PhyA-based photoswitch) system ( 176 ) have been created to achieve light-induced ERK activation in Drosophila and mammals.…”

Section: Application Of Optogenetics In Physiological Processesmentioning

confidence: 99%

Optophysiology: Illuminating cell physiology with optogenetics

Tan

Huang

et al. 2022

Physiological Reviews

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Optogenetics combines light and genetics to enable precise control of living cells, tissues, and organisms with tailored functions. Optogenetics has the advantages of noninvasiveness, rapid responsiveness, tunable reversibility, and superior spatiotemporal resolution. Following the initial discovery of microbial opsins as light-actuated ion channels, a plethora of naturally occurring or engineered photoreceptors or photosensitive domains that respond to light at varying wavelengths has ushered in the next chapter of optogenetics. Through protein engineering and synthetic biology approaches, genetically-encoded photoswitches can be modularly engineered into protein scaffolds or host cells to control a myriad of biological processes, as well as to enable behavioral control and disease intervention in vivo. Here, we summarize these optogenetic tools on the basis of their fundamental photochemical properties to better inform the chemical basis and design principles. We also highlight exemplary applications of opsin-free optogenetics in dissecting cellular physiology (designated "optophysiology"), and describe the current progress, as well as future trends, in wireless optogenetics, which enables remote interrogation of physiological processes with minimal invasiveness. This review is anticipated to spark novel thoughts on engineering next-generation optogenetic tools and devices that promise to accelerate both basic and translational studies.

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“…To tackle the above‐mentioned roadblocks, the Ye group continued to develop a red‐shifted optogenetic devices, termed as REDMAP for red/far‐red light‐mediated and miniaturized PhyA (ΔPhyA)‐based photoswitch 9 (Figure 1A,B ). REDMAP is composed of the photosensory module of Arabidopsis photoreceptor PhyA and the shuttle protein far‐red elongated hypocotyl 1 (FHY1).…”

Section: Redmap As a Miniature Optogenetic Systemmentioning

confidence: 99%

“…Compared to the PhyB/PIF combination, the REDMAP system has a compact size of only ∼3.2 kb, and therefore, can be packaged into clinically approved AAV vectors for efficient delivery into rodent livers. 9 The REDMAP components can be engineered into the yeast Gal4 DNA‐binding domain and the artificial transactivator VP64 (Figure 1C ), thereby achieving potent transcriptional activation (>150‐fold) with fast kinetics (∼1 s for activation) and mild light dose (1 mW/cm 2 ). In living mammals, REDMAP was shown to enable red light‐inducible expression of insulin in type 1 diabetic mice and rat, as well as transgene expression in rabbits.…”

Section: Redmap As a Miniature Optogenetic Systemmentioning

confidence: 99%