General Method for Regulating Protein Stability with Light

Bonger, Kimberly M.; Rakhit, Rishi; Payumo, Alexander Y.; Chen, James; Wandless, Thomas J.

doi:10.1021/cb400755b

Cited by 135 publications

(153 citation statements)

References 20 publications

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“…Three are based on photosensitive plant proteins (cryptochromes 9–11 , light-oxygen-voltage (LOV) domains 12–15 and phytochromes 16–18 ), and one is based on the fluorescent protein Dronpa 19 , which was isolated from the coral Pectiniidae 20 . Other recent publications discuss the use of optogenetic proteins that manipulate specific signalling events, such as those that regulate neuronal excitability 4,21 , cyclic nucleotides 22,23 and heterotrimeric G protein signalling 24,25 , or proteins that are irreversibly activated 26–28 or inactivated 29 by light.…”

Section: Overview Of Optogenetic Systemsmentioning

confidence: 99%

Illuminating cell signalling with optogenetic tools

Tischer

Weiner

2014

Nat Rev Mol Cell Biol

317

282

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The light-based control of ion channels has been transformative for the neurosciences, but the optogenetic toolkit does not stop there. An expanding number of proteins and cellular functions have been shown to be controlled by light, and the practical considerations in deciding between reversible optogenetic systems (such as systems that use light-oxygen-voltage domains, phytochrome proteins, cryptochrome proteins and the fluorescent protein Dronpa) are well defined. The field is moving beyond proof of concept to answering real biological questions, such as how cell signalling is regulated in space and time, that were difficult or impossible to address with previous tools.

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Section: Overview Of Optogenetic Systemsmentioning

confidence: 99%

Illuminating cell signalling with optogenetic tools

Tischer

Weiner

2014

Nat Rev Mol Cell Biol

317

282

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“…This toolbox now offers the possibility to photocontrol a variety of cellular protein functions from chromatin modification to DNA transcription [20][21][22][23][24][25][26] or recombination 23 , protein translocation 19,22,27,28 , enzymatic activity [29][30][31] , cell morphology 22,32 , signaling pathways 7,27,30,31 and protein degradation 33,34 (Table 1). Recent works have focused on improving the response timescale and the signal-to-noise ratio and have expanded the spectrum of available wavelengths ( Table 1), making multiplexing possible (Box 1 and Fig.…”

Section: Genetically Encodable Photoactuatorsmentioning

confidence: 99%

How to control proteins with light in living systems

et al. 2014

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The possibility offered by photocontrolling the activity of biomolecules in vivo while recording physiological parameters is opening up new opportunities for the study of physiological processes at the single-cell level in a living organism. For the last decade, such tools have been mainly used in neuroscience, and their application in freely moving animals has revolutionized this field. New photochemical approaches enable the control of various cellular processes by manipulating a wide range of protein functions in a noninvasive way and with unprecedented spatiotemporal resolution. We are at a pivotal moment where biologists can adapt these cutting-edge technologies to their system of study. This user-oriented review presents the state of the art and highlights technical issues to be resolved in the near future for wide and easy use of these powerful approaches.

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“…While we already observed strong inhibition of reporter expression from the VP16-and VP64-based systems when Med25VBD was transfected in equal amounts to the other components (10-fold and 20-fold respectively), the p65-based system was barely repressed 2-fold at a 5-fold Having confirmed that Med25VBD can be used to potently and selectively repress VP64-dependent transgene expression, we embarked upon the engineering of a light-controlled VP64 inhibitor. To this end, we turned to the recently-published B-LID (blue light-inducible degradation) domain (Bonger et al 2014). This optogenetic protein degradation tool is based on the LOV2 domain from Avena sativa phototropin 1 that has been modified by adding an RRRG-degradation signal to its J-helix.…”

Section: Acc E P Ted P R E P R I Ntmentioning

confidence: 99%

An optogenetic upgrade for the Tet‐OFF system

Müller

Zurbriggen

Weber

2015

Biotech & Bioengineering

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The rapid development of mammalian optogenetics has produced an expanding number of gene switches that can be controlled with the unprecedented spatiotemporal resolution of light. However, in the "pre-optogenetic" era many networks, cell lines and transgenic organisms have been engineered that rely on chemically-inducible transgene expression systems but would benefit from the advantages of the traceless inducer light. To open the possibility for the effortless upgrade of such systems from chemical inducers to light, we capitalized on the specific Med25VBD inhibitor of the VP16/VP64 transactivation domain. In a first step, we demonstrated the efficiency and selectivity of Med25VBD in the inhibition of VP16/VP64-based transgene expression systems. Then, we fused the inhibitor to the blue light-responsive B-LID degron and optimized the performance of this construct with regard to the number of Med25VBD repeats. This approach resulted in an optogenetic upgrade of the popular Tet-OFF (TetR-VP64, tetO7 -PhCMVmin ) system that allows tunable, blue light-inducible transgene expression in HEK-293T cells.

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General Method for Regulating Protein Stability with Light

Cited by 135 publications

References 20 publications

Illuminating cell signalling with optogenetic tools

Illuminating cell signalling with optogenetic tools

How to control proteins with light in living systems

An optogenetic upgrade for the Tet‐OFF system

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