Near‐Infrared Light‐Controlled Gene Expression and Protein Targeting in Neurons and Non‐neuronal Cells

Redchuk, Taras A.; Karasev, Maksim M.; Omelina, Evgeniya S.; Verkhusha, Vladislav V.

doi:10.1002/cbic.201700642

Cited by 24 publications

(24 citation statements)

References 34 publications

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“…It has been shown that a two-component FR–NIR optogenetic system based on the heterodimerization of a RpBphP1 bacterial phytochrome with its binding partner RpPpsR2 is spectrally compatible with LOV-based optogenetic tools, enabling simultaneous regulation of several processes in a cell 30,31 . Our current results indicate that the FR–NIR light sensitivity of the DrBphP-based opto-kinases also allows their crosstalk-free combination with the visible-light optogenetic tools and fluorescent proteins.…”

Section: Discussionmentioning

confidence: 99%

Neurotrophin receptor tyrosine kinases regulated with near-infrared light

et al. 2019

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Optical control over the activity of receptor tyrosine kinases (RTKs) provides an efficient way to reversibly and non-invasively map their functions. We combined catalytic domains of Trk (tropomyosin receptor kinase) family of RTKs, naturally activated by neurotrophins, with photosensory core module of DrBphP bacterial phytochrome to develop opto-kinases, termed Dr-TrkA and Dr-TrkB, reversibly switchable on and off with near-infrared and far-red light. We validated Dr-Trk ability to reversibly light-control several RTK pathways, calcium level, and demonstrated that their activation triggers canonical Trk signaling. Dr-TrkA induced apoptosis in neuroblastoma and glioblastoma, but not in other cell types. Absence of spectral crosstalk between Dr-Trks and blue-light-activatable LOV-domain-based translocation system enabled intracellular targeting of Dr-TrkA independently of its activation, additionally modulating Trk signaling. Dr-Trks have several superior characteristics that make them the opto-kinases of choice for regulation of RTK signaling: high activation range, fast and reversible photoswitching, and multiplexing with visible-light-controllable optogenetic tools.

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Section: Discussionmentioning

confidence: 99%

Neurotrophin receptor tyrosine kinases regulated with near-infrared light

et al. 2019

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“…Combining optogenetic tools controlled by light of different wavelengths enabled tridirectional protein targeting 27 , which may be used to control multifunctional proteins or to increase dynamic range of bidirectional localizers, as discussed 31 . A NIR-blue-light inducible shuttle (iRIS) construct was shown to be effective for dualwavelength-controlled exogenous protein relocalization 16,19 . In this system, the QPAS1 protein is fused to a blue-light-sensing AsLOV2-based nuclear localization controller.…”

Section: Resultsmentioning

confidence: 99%

“…Blue-light-mediated genome editing was reported recently 17 . The non-opsin optogenetic tools are used in cultured cells of various origin 18 , in primary cell cultures 19 and in vivo 20 . Although the optogenetic systems to regulate protein-protein interactions demonstrated high spatiotemporal precision and large dynamic range of responces 21,22 , they usually control functions of the overexpressed exogenous proteins.…”

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confidence: 99%

Optogenetic regulation of endogenous proteins

et al. 2020

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Techniques of protein regulation, such as conditional gene expression, RNA interference, knock-in and knockout , lack sufficient spatiotemporal accuracy, while optogenetic tools suffer from non-physiological response due to overexpression artifacts. Here we present a near-infrared light-activatable optogenetic system, which combines the specificity and orthogonality of intrabodies with the spatiotemporal precision of optogenetics. We engineer optically-controlled intrabodies to regulate genomically expressed protein targets and validate the possibility to further multiplex protein regulation via dual-wavelength optogenetic control. We apply this system to regulate cytoskeletal and enzymatic functions of two nontagged endogenous proteins, actin and RAS GTPase, involved in complex functional networks sensitive to perturbations. The optogenetically-enhanced intrabodies allow fast and reversible regulation of both proteins, as well as simultaneous monitoring of RAS signaling with visiblelight biosensors, enabling all-optical approach. Growing number of intrabodies should make their incorporation into optogenetic tools the versatile technology to regulate endogenous targets.

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“…Upon illumination, activated BphP1 binds to PpsR2 and releases the promoter under its repression. UV light is harmful to cells while near-infrared signals can penetrate tissues (Chen et al, 2018 ), which enables applications of the BphP1/PpsR2 system in mammalian cells (Kaberniuk et al, 2016 ; Redchuk et al, 2017 , 2018b ).…”

Section: Light Sensors In Bacteriamentioning

confidence: 99%

Programming Bacteria With Light—Sensors and Applications in Synthetic Biology

et al. 2018

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Photo-receptors are widely present in both prokaryotic and eukaryotic cells, which serves as the foundation of tuning cell behaviors with light. While practices in eukaryotic cells have been relatively established, trials in bacterial cells have only been emerging in the past few years. A number of light sensors have been engineered in bacteria cells and most of them fall into the categories of two-component and one-component systems. Such a sensor toolbox has enabled practices in controlling synthetic circuits at the level of transcription and protein activity which is a major topic in synthetic biology, according to the central dogma. Additionally, engineered light sensors and practices of tuning synthetic circuits have served as a foundation for achieving light based real-time feedback control. Here, we review programming bacteria cells with light, introducing engineered light sensors in bacteria and their applications, including tuning synthetic circuits and achieving feedback controls over microbial cell culture.

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Near‐Infrared Light‐Controlled Gene Expression and Protein Targeting in Neurons and Non‐neuronal Cells

Cited by 24 publications

References 34 publications

Neurotrophin receptor tyrosine kinases regulated with near-infrared light

Neurotrophin receptor tyrosine kinases regulated with near-infrared light

Optogenetic regulation of endogenous proteins

Programming Bacteria With Light—Sensors and Applications in Synthetic Biology

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