Shedding light on current trends in molecular optogenetics

Fischer, Alexandra; Kramer, Markus M.; Radziwill, Gerald; Weber, Wilfried

doi:10.1016/j.cbpa.2022.102196

Cited by 15 publications

(19 citation statements)

References 64 publications

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“…Here, we review the current state and recent developments of light-regulated gene expression in prokaryotes. Since the arrival of the initial optogenetic setups for gene expression in bacteria (Levskaya et al, 2005;Möglich et al, 2009;Tabor et al, 2011;Ohlendorf et al, 2012), many more systems were advanced (Baumschlager and Khammash, 2021;Fischer et al, 2022;Hoffman et al, 2022;Lindner and Diepold, 2022;Mazraeh and Di Ventura, 2022;Reshetnikov et al, 2022). In this article, we first recapitulate fundamental aspects of photoreceptors and optogenetics as they pertain to lightregulated gene expression.…”

Section: Introductionmentioning

confidence: 99%

Light-regulated gene expression in Bacteria: Fundamentals, advances, and perspectives

Ohlendorf

Möglich²

2022

Front. Bioeng. Biotechnol.

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Numerous photoreceptors and genetic circuits emerged over the past two decades and now enable the light-dependent i.e., optogenetic, regulation of gene expression in bacteria. Prompted by light cues in the near-ultraviolet to near-infrared region of the electromagnetic spectrum, gene expression can be up- or downregulated stringently, reversibly, non-invasively, and with precision in space and time. Here, we survey the underlying principles, available options, and prominent examples of optogenetically regulated gene expression in bacteria. While transcription initiation and elongation remain most important for optogenetic intervention, other processes e.g., translation and downstream events, were also rendered light-dependent. The optogenetic control of bacterial expression predominantly employs but three fundamental strategies: light-sensitive two-component systems, oligomerization reactions, and second-messenger signaling. Certain optogenetic circuits moved beyond the proof-of-principle and stood the test of practice. They enable unprecedented applications in three major areas. First, light-dependent expression underpins novel concepts and strategies for enhanced yields in microbial production processes. Second, light-responsive bacteria can be optogenetically stimulated while residing within the bodies of animals, thus prompting the secretion of compounds that grant health benefits to the animal host. Third, optogenetics allows the generation of precisely structured, novel biomaterials. These applications jointly testify to the maturity of the optogenetic approach and serve as blueprints bound to inspire and template innovative use cases of light-regulated gene expression in bacteria. Researchers pursuing these lines can choose from an ever-growing, versatile, and efficient toolkit of optogenetic circuits.

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Section: Introductionmentioning

confidence: 99%

Light-regulated gene expression in Bacteria: Fundamentals, advances, and perspectives

Ohlendorf

Möglich²

2022

Front. Bioeng. Biotechnol.

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“…Manipulation of the ligand concentration, the affinity and the half-life of the ligand-interaction, the timing of when the ligand binds or not and the location where the ligand binds, is important to interrogate how cells react to a ligand under changing conditions. One novel technology to temporally modulate the quantity, kinetics and localization of a ligand binding to a receptor, is optogenetics (Fischer et al, 2022;Sánchez and Tampé, 2022).…”

Section: Introductionmentioning

confidence: 99%

“…Modulation of the light-intensity can be used to control the amount of X and Y that form the X-Y complex or to control the affinity and half-life of the interaction (Tischer and Weiner, 2019;Yousefi et al, 2019). The timing and localisation of the light illumination can be used to regulate the X-Y interaction with an unprecedentedly high spatio-temporal resolution over several orders of magnitude, ranging from sub-seconds to days and from micrometer to meter (Kolar et al, 2018;Fischer et al, 2022).…”

Section: Introductionmentioning

confidence: 99%

Opto-APC: Engineering of cells that display phytochrome B on their surface for optogenetic studies of cell-cell interactions

Russ

Ehret²,

Hörner³

et al. 2023

Front. Mol. Biosci.

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The kinetics of a ligand-receptor interaction determine the responses of the receptor-expressing cell. One approach to experimentally and reversibly change this kinetics on demand is optogenetics. We have previously developed a system in which the interaction of a modified receptor with an engineered ligand can be controlled by light. In this system the ligand is a soluble Phytochrome B (PhyB) tetramer and the receptor is fused to a mutated PhyB-interacting factor (PIFS). However, often the natural ligand is not soluble, but expressed as a membrane protein on another cell. This allows ligand-receptor interactions in two dimensions. Here, we developed a strategy to generate cells that display PhyB as a membrane-bound protein by expressing the SpyCatcher fused to a transmembrane domain in HEK-293T cells and covalently coupling purified PhyB-SpyTag to these cells. As proof-of-principle, we use Jurkat T cells that express a GFP-PIFS-T cell receptor and show that these cells can be stimulated by the PhyB-coupled HEK-293T cells in a light dependent manner. Thus, we call the PhyB-coupled cells opto-antigen presenting cells (opto-APCs). Our work expands the toolbox of optogenetic technologies, allowing two-dimensional ligand-receptor interactions to be controlled by light.

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“…The optimal stimulus for inducing such alterations is light, as it can be applied in a reversible, adjustable, and local manner at excellent resolution. [ 23 ] In order to translate optical input into changes in the hydrogels’ mechanical properties, chemical or biological photo‐responsive moieties are required. For such applications, biological photoreceptors show highly suitable features as they are inherently functional in a biological background, are responsive to the biocompatible light spectrum and intensities, and show high specificity, even in a complex biochemical environment.…”

Section: Introductionmentioning

confidence: 99%

A Photoreceptor‐Based Hydrogel with Red Light‐Responsive Reversible Sol‐Gel Transition as Transient Cellular Matrix

Hörner

Becker

Bohnert

et al. 2023

Adv Materials Technologies

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Hydrogels with adjustable mechanical properties have been engineered as matrices for mammalian cells and allow the dynamic, mechano‐responsive manipulation of cell fate and function. Recent research yields hydrogels, where biological photoreceptors translated optical signals into a reversible and adjustable change in hydrogel mechanics. While their initial application provides important insights into mechanobiology, broader implementation is limited by a small dynamic range of addressable stiffness. Herein, this limitation is overcome by developing a photoreceptor‐based hydrogel with reversibly adjustable stiffness from ≈800 Pa to the sol state. The hydrogel is based on star‐shaped polyethylene glycol, functionalized with the red/far‐red light photoreceptor phytochrome B (PhyB), or phytochrome‐interacting factor 6 (PIF6). Upon illumination with red light, PhyB heterodimerizes with PIF6, thus crosslinking the polymers and resulting in gelation. However, upon illumination with far‐red light, the proteins dissociate and trigger a complete gel‐to‐sol transition. The hydrogel's light‐responsive mechanical properties are comprehensively characterized and it is applied as a reversible extracellular matrix for the spatiotemporally controlled deposition of mammalian cells within a microfluidic chip. It is anticipated that this technology will open new avenues for the site‐ and time‐specific positioning of cells and will contribute to overcome spatial restrictions.

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Shedding light on current trends in molecular optogenetics

Cited by 15 publications

References 64 publications

Light-regulated gene expression in Bacteria: Fundamentals, advances, and perspectives

Light-regulated gene expression in Bacteria: Fundamentals, advances, and perspectives

Opto-APC: Engineering of cells that display phytochrome B on their surface for optogenetic studies of cell-cell interactions

A Photoreceptor‐Based Hydrogel with Red Light‐Responsive Reversible Sol‐Gel Transition as Transient Cellular Matrix

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