Lights, cytoskeleton, action: Optogenetic control of cell dynamics

Wittmann, Torsten; Dema, Alessandro; Haren, Jeffrey van

doi:10.1016/j.ceb.2020.03.003

Cited by 38 publications

(31 citation statements)

References 81 publications

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“…A possible application for the high spatial control of optogenetics is the targeting of subcellular compartments of the cell. For an in-depth review of optogenetic control of cell dynamics and subcellular applications see the recently published reviews [ 91 , 92 ], An example is the optogenetic TULIP system based on the LOV2-domain that was applied for local activation and recruitment of the RhoA GTPase to the membrane in epithelial cells ( Figure 6 A). This optogenetic tool enables a short and spatially controlled activation of RhoA.…”

Section: Control Of Subcellular Localization and Spatial Resolution Of Signaling Processesmentioning

confidence: 99%

Optogenetic Approaches for the Spatiotemporal Control of Signal Transduction Pathways

Kramer

Lataster

Weber

et al. 2021

IJMS

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Biological signals are sensed by their respective receptors and are transduced and processed by a sophisticated intracellular signaling network leading to a signal-specific cellular response. Thereby, the response to the signal depends on the strength, the frequency, and the duration of the stimulus as well as on the subcellular signal progression. Optogenetic tools are based on genetically encoded light-sensing proteins facilitating the precise spatiotemporal control of signal transduction pathways and cell fate decisions in the absence of natural ligands. In this review, we provide an overview of optogenetic approaches connecting light-regulated protein-protein interaction or caging/uncaging events with steering the function of signaling proteins. We briefly discuss the most common optogenetic switches and their mode of action. The main part deals with the engineering and application of optogenetic tools for the control of transmembrane receptors including receptor tyrosine kinases, the T cell receptor and integrins, and their effector proteins. We also address the hallmarks of optogenetics, the spatial and temporal control of signaling events.

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Section: Control Of Subcellular Localization and Spatial Resolution Of Signaling Processesmentioning

confidence: 99%

Optogenetic Approaches for the Spatiotemporal Control of Signal Transduction Pathways

Kramer

Lataster

Weber

et al. 2021

IJMS

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“…Photouncageable antimitotics were the first to be developed and have achieved some key applications [18,19]; however, they faced several limitations and are no longer commercially available. Optogenetic approaches to manipulate MTs have only been reported recently [20,21], and will no doubt continue to develop [22].…”

Section: Notesmentioning

confidence: 99%

Photocontrolling Microtubule Dynamics with Photoswitchable Chemical Reagents

Thorn‐Seshold

Meiring²

2021

Preprint

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Microtubule dynamics can be inhibited with sub-second temporal resolution and cellular-scale spatial resolution, by using precise illuminations to optically pattern where and when photoswitchable microtubule-inhibiting chemical reagents exert their latent bioactivity. The recently-available reagents (SBTub, PST, STEpo, AzTax, PHTub) now enable researchers to use light to reversibly modulate microtubule-dependent processes in eukaryotes, in 2D and 3D cell culture as well as in vivo, across a variety of model organisms: with applications in fields from cargo transport to cell migration, cell division, and embryonic development. However, a wide knowledge gap has remained in the literature, which has blocked further translation of these and many other classes of photopharmaceuticals. No generally-applicable procedures or workflows to establish biological assays using photopharmaceuticals have been published. Accordingly, the rate of adoption of photopharmaceutical tools in the broader chemical biology community (beyond the original chemical developers of the tools) has remained very low. Vital information about assay benchmarking for photoconversion, testing for isomer solubility, proving the retention of mechanism of action, estimating the limits of phototoxicity etc has either simply not been formalised in the literature, or has remained buried in diverse reports without being unified and codified for an audience beyond that of synthetic organic chemists. Here we have developed a robust four-step assay establishment procedure to optimise assay parameters for achieving reliable photocontrol over microtubule dynamics, that is applicable to diverse families of photoswitchable inhibitors. This procedure also controls for these common sources of irreproducibility and includes numerous troubleshooting steps. We also collect together the relevant information for non-chemist "users" such as microscopists and biologists, to introduce the theory of small molecule photoswitching; the unique features, usage requirements, and limitations that photoswitchable chemical reagents have; and the specific performance features of the major classes of photoswitchable microtubule inhibitors that are currently available; to highlight their properties that suit them to different applications. The generally-applicable workflows that we present allow establishing cellular assays optically controlling microtubule dynamics in a temporally reversible fashion with spatial specificity down to a single selected cell within a field of view. These workflows and methods also equip the reader to tackle advanced uses of photoswitchable chemical reagents for general protein targets, in 3D culture and in vivo, and can represent an important bridge to reach the high-value biological applications that photopharmacology can promise.

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“…Unlike optogenetics, photopharmaceuticals do not need genetic engineering, so they transition easily between biological models; and they can target proteins which do not allow functional photocontrol by fusion constructs, such as the cytoskeletal scaffolds tubulin and actin [3, 8, 9] . Since the spatiotemporally precise orchestration of cytoskeleton structure and dynamics is critically important to hundreds of biological processes, cytoskeleton photopharmaceuticals have emerged as a valuable goal of research in recent years [10, 11] . They hold promise for basic research into anisotropic processes, for example, in intracellular transport, mechanostasis and cell motility, as well as for applied research on, for example, anti‐invasive strategies in cancer therapy [10, 12, 13] …”

Section: Introductionmentioning

confidence: 99%

Pyrrole Hemithioindigo Antimitotics with Near‐Quantitative Bidirectional Photoswitching that Photocontrol Cellular Microtubule Dynamics with Single‐Cell Precision**

et al. 2021

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We report the first cellular application of the emerging near‐quantitative photoswitch pyrrole hemithioindigo, by rationally designing photopharmaceutical PHTub inhibitors of the cytoskeletal protein tubulin. PHTubs allow simultaneous visible‐light imaging and photoswitching in live cells, delivering cell‐precise photomodulation of microtubule dynamics, and photocontrol over cell cycle progression and cell death. This is the first acute use of a hemithioindigo photopharmaceutical for high‐spatiotemporal‐resolution biological control in live cells. It additionally demonstrates the utility of near‐quantitative photoswitches, by enabling a dark‐active design to overcome residual background activity during cellular photopatterning. This work opens up new horizons for high‐precision microtubule research using PHTubs and shows the cellular applicability of pyrrole hemithioindigo as a valuable scaffold for photocontrol of a range of other biological targets.

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Lights, cytoskeleton, action: Optogenetic control of cell dynamics

Cited by 38 publications

References 81 publications

Optogenetic Approaches for the Spatiotemporal Control of Signal Transduction Pathways

Optogenetic Approaches for the Spatiotemporal Control of Signal Transduction Pathways

Photocontrolling Microtubule Dynamics with Photoswitchable Chemical Reagents

Pyrrole Hemithioindigo Antimitotics with Near‐Quantitative Bidirectional Photoswitching that Photocontrol Cellular Microtubule Dynamics with Single‐Cell Precision**

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