FLIRT: fast local infrared thermogenetics for subcellular control of protein function

Hirsch, Sophia M.; Sundaramoorthy, Sriramkumar; Davies, Tim; Zhuravlev, Yelena; Waters, Jennifer; Shirasu-Hiza, Mimi; Dumont, Julien; Canman, Julie C.

doi:10.1038/s41592-018-0168-y

Cited by 27 publications

(25 citation statements)

References 31 publications

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“…Our data imply that cell-cycle timings are controlled in a cell-autonomous manner, with no significant contribution from cell-to-cell communication. These results open multiple future directions, including the exploration of cell-cycle-timing control at later developmental stages and further investigations of the developmental consequence of perturbing cell-cycle times, as well as additional thermogenetic control (27,(32)(33)(34). In vivo nanoscale NV thermometry allows for the long-term readout of temperature at subcellular levels without photo-bleaching (10,35) improvements utilizing magnetic-criticality-enhanced measurements (36).…”

Section: Discussionmentioning

confidence: 93%

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Probing and manipulating embryogenesis via nanoscale thermometry and temperature control

Choi

Zhou

Landig

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

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Understanding the coordination of cell-division timing is one of the outstanding questions in the field of developmental biology. One active control parameter of the cell-cycle duration is temperature, as it can accelerate or decelerate the rate of biochemical reactions. However, controlled experiments at the cellular scale are challenging, due to the limited availability of biocompatible temperature sensors, as well as the lack of practical methods to systematically control local temperatures and cellular dynamics. Here, we demonstrate a method to probe and control the cell-division timing inCaenorhabditis elegansembryos using a combination of local laser heating and nanoscale thermometry. Local infrared laser illumination produces a temperature gradient across the embryo, which is precisely measured by in vivo nanoscale thermometry using quantum defects in nanodiamonds. These techniques enable selective, controlled acceleration of the cell divisions, even enabling an inversion of division order at the two-cell stage. Our data suggest that the cell-cycle timing asynchrony of the early embryonic development inC. elegansis determined independently by individual cells rather than via cell-to-cell communication. Our method can be used to control the development of multicellular organisms and to provide insights into the regulation of cell-division timings as a consequence of local perturbations.

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Section: Discussionmentioning

confidence: 93%

“…2 A, Left). For local heating, we employed an infrared (IR) laser at a wavelength of 1,480 nm (26,27), selectively focused on targeted cells with a beam waist of ∼2 µm. To accurately determine the temperature distribution under local laser illumination experimentally, it is crucial to monitor the temperature in vivo.…”

Section: Methodsmentioning

confidence: 99%

Probing and manipulating embryogenesis via nanoscale thermometry and temperature control

Choi

Zhou

Landig

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

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“…FLUCS is localized, directed, dynamic, physically tractable, physiological, and strictly non-invasive, as confirmed by assays of development and sensitive stress-response measurements. The temperature amplitudes required are usually in the order of only 1 C-3 C, which is comparable to or lower than the heating observed in thermo-genetics (Hirsch et al, 2018).…”

Section: Physical Perturbations To Dissect the Functional Role Of Difmentioning

confidence: 85%

“…Specifically, spatially localized activation of light-inducible molecular machinery ( Figure 2G) can be used to induce and guide developmental programs such as epithelial folding in fly embryos (Izquierdo et al, 2018). Interestingly, a minimally invasive inactivation of temperature-sensitive motor proteins ( Figure 2H) has also recently been employed in embryos and adult worms (Hirsch et al, 2018).…”

Section: Physical Perturbations To Dissect the Functional Role Of Difmentioning

confidence: 99%

Probing the Functional Role of Physical Motion in Development

Kreysing

2019

Developmental Cell

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“…Strain JCC596 [19], kindly provided by Sophia Hirsch and Julie Canman, includes zuls178 , which marks all nuclei with HIS-72::GFP, and stls10138 , which marks pharyngeal cells with mCherry:HistoneH1 and can be used assess GLP-1/Notch activity at the four-cell stage: it is expressed in approximately 25% of the cells in late-stage embryos when GLP-1 has been activated in ABp, and in approximately 50% of cells when Notch activation in ABp has not occurred.…”

Section: Star Methodsmentioning

confidence: 99%

The C. elegans Notch proteins LIN-12 and GLP-1 are tuned to lower force thresholds for activation than Drosophila Notch

García-Díaz

et al. 2021

Preprint

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The conserved transmembrane receptor Notch mediates cell fate decisions in all animals. In the absence of ligand, a Negative Regulatory Region (NRR) in the Notch ectodomain adopts an autoinhibited confirmation, masking an ADAM protease cleavage site [1, 2]; ligand binding makes the cleavage site accessible, leading to shedding of the Notch ectodomain as the first step of signal transduction [3, 4]. In Drosophila and vertebrates, the ligands are all single-pass transmembrane Delta/Serrate/LAG-2 (DSL) proteins; the endocytic adaptor Epsin binds to the ubiquitinated intracellular domain, and the resulting Clathrin-mediated endocytosis exerts a "pulling force" that exposes the cleavage site in the NRR [4-6]. However, in C. elegans, the presence of natural secreted DSL proteins [7] and other observations suggested that Epsin-mediated endocytosis may not be required to activate the Notch proteins LIN-12 and GLP-1. Here, we confirm that neither Epsin nor the cytosolic domains of DSL proteins are required for Notch signaling in C. elegans. Furthermore, we provide evidence that the NRRs of LIN-12 and GLP-1 are tuned to a lower force level than the NRR of Drosophila Notch. Finally, we show that adding a Leucine "plug" that occludes the cleavage site in vertebrate and Drosophila Notch proteins but is absent in the C. elegans Notch proteins [1, 2] renders the LIN-12 and GLP-1 NRRs dependent on Epsin-mediated ligand endocytosis, indicating that greater force is now required to expose the cleavage site. Thus, the NRRs of LIN-12 and GLP-1 appear to be tuned to a lower force threshold, accounting for the different requirements for signaling in C. elegans.

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FLIRT: fast local infrared thermogenetics for subcellular control of protein function

Cited by 27 publications

References 31 publications

Probing and manipulating embryogenesis via nanoscale thermometry and temperature control

Probing and manipulating embryogenesis via nanoscale thermometry and temperature control

Probing the Functional Role of Physical Motion in Development

The C. elegans Notch proteins LIN-12 and GLP-1 are tuned to lower force thresholds for activation than Drosophila Notch

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