Optogenetic control of the Bicoid morphogen reveals fast and slow modes of gap gene regulation

Singh, Annette; Wu, Ping; Ryabichko, Sergey; Raimundo, João; Swan, M.K.; Wieschaus, Eric; Gregor, Thomas; Toettcher, Jared E.

doi:10.1016/j.celrep.2022.110543

Cited by 34 publications

(22 citation statements)

References 67 publications

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“…Notably, optogenetics ( Figure 3 c) – the capability to control biochemical reactions by light – is proving to be extremely powerful in probing biochemical networks [ 69 , 70 , 71 , 72 ]. For example, Wilson et al.…”

Section: Perspectives and Conclusionmentioning

confidence: 99%

Signalling dynamics, cell decisions, and homeostatic control in health and disease

Valls

Esposito

2022

Current Opinion in Cell Biology

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Section: Perspectives and Conclusionmentioning

confidence: 99%

Signalling dynamics, cell decisions, and homeostatic control in health and disease

Valls

Esposito

2022

Current Opinion in Cell Biology

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“…DMDs are extensively used as spatial light modulators in commercial projection systems ( Dudley et al, 2003 ), and here in these platforms DMDs facilitated projecting user-defined mask images onto the microscope sample plane where the target cells are present ( Figure 3A ). In the last decade, several improved microscope-coupled DMD or LCD projector setups have been designed and demonstrated in cellular optogenetic control applications ( Zhu et al, 2012 ; Chait et al, 2017 ; Rullan et al, 2018 ; Kumar et al, 2021 ; Fox et al, 2022 ; Singh et al, 2021 ). In most of these platforms, a DMD ( Rullan et al, 2018 ) or LCD ( Chait et al, 2017 ) projector, along with additional optical elements like lenses, filters, etc ., is placed at the episcopic illumination port of the microscope.…”

Section: Low-volume Culture Platformsmentioning

confidence: 99%

Platforms for Optogenetic Stimulation and Feedback Control

Kumar

Khammash²

2022

Front. Bioeng. Biotechnol.

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Harnessing the potential of optogenetics in biology requires methodologies from different disciplines ranging from biology, to mechatronics engineering, to control engineering. Light stimulation of a synthetic optogenetic construct in a given biological species can only be achieved via a suitable light stimulation platform. Emerging optogenetic applications entail a consistent, reproducible, and regulated delivery of light adapted to the application requirement. In this review, we explore the evolution of light-induction hardware-software platforms from simple illumination set-ups to sophisticated microscopy, microtiter plate and bioreactor designs, and discuss their respective advantages and disadvantages. Here, we examine design approaches followed in performing optogenetic experiments spanning different cell types and culture volumes, with induction capabilities ranging from single cell stimulation to entire cell culture illumination. The development of automated measurement and stimulation schemes on these platforms has enabled researchers to implement various in silico feedback control strategies to achieve computer-controlled living systems—a theme we briefly discuss in the last part of this review.

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“…LEXY has been shown to work robustly in many mammalian cell lines, 4,5 in Xenopus epithelial cells and mouse zygotes, 6 and Drosophila. 7,8 In a recent study, focused on understanding how the transcription factor Twist functions in Drosophila embryos, the LOV domain within LEXY was mutated, and two variants, iLEXYi and iLEXYs, were obtained. 9 The authors chose these names to indicate that these variants are an improvement to the original LEXY, allowing for a more complete depletion of Twist from the nucleus and the usage of lower light intensities or longer intervals between light pulses.…”

mentioning

confidence: 99%

“…LEXY has been shown to work robustly in many mammalian cell lines, 4 , 5 in Xenopus epithelial cells and mouse zygotes, 6 and Drosophila . 7 , 8 …”

mentioning

confidence: 99%

“…Quickly after blue light illumination is ceased, the POI accumulates back into the nucleus because the As LOV2 domain reverts back to the dark state within about 80 s making the NES again inaccessible and letting the NLS take over. LEXY has been shown to work robustly in many mammalian cell lines, , in Xenopus epithelial cells and mouse zygotes, and Drosophila . , …”

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confidence: 99%

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Analysis of Slow-Cycling Variants of the Light-Inducible Nuclear Protein Export System LEXY in Mammalian Cells

et al. 2022

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The optogenetic tool LEXY consists of the second light oxygen voltage (LOV) domain of Avena sativa phototropin 1 mutated to contain a nuclear export signal. It allows exporting from the nucleus with blue light proteins of interest (POIs) genetically fused to it. Mutations slowing the dark recovery rate of the LOV domain within LEXY were recently shown to allow for better depletion of some POIs from the nucleus in Drosophila embryos and for the usage of low light illumination regimes. We investigated these variants in mammalian cells and found they increase the cytoplasmic localization of the proteins we tested after illumination, but also during the dark phases, which corresponds to higher leakiness of the system. These data suggest that, when aiming to sequester into the nucleus a protein with a cytoplasmic function, the original LEXY is preferable. The iLEXY variants are, instead, advantageous when wanting to deplete the nucleus of the POI as much as possible.

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Optogenetic control of the Bicoid morphogen reveals fast and slow modes of gap gene regulation

Cited by 34 publications

References 67 publications

Signalling dynamics, cell decisions, and homeostatic control in health and disease

Signalling dynamics, cell decisions, and homeostatic control in health and disease

Platforms for Optogenetic Stimulation and Feedback Control

Analysis of Slow-Cycling Variants of the Light-Inducible Nuclear Protein Export System LEXY in Mammalian Cells

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