Abstract:We previously developed the Magnet system, which consists of two distinct Vivid protein variants, one positively and one negatively charged, designated the positive Magnet (pMag) and negative Magnet (nMag), respectively. These two proteins bind to each other through electrostatic interactions, preventing unwanted homodimerization and providing selective light-induced heterodimerization. The Magnet system enables the manipulation of cellular functions such as protein-protein interactions and genome editing, alt… Show more
“…The Magnet system (pMag and nMag pair) derived from Vivid has the advantage of small size (151 aa), allowing rapid heterodimerization reversibility (half‐life is 6.8 s) . Additionally, a CAD‐Magnet system with increased avidity between pMag and nMag was developed and used to induce membrane ruffling . There are two reports that the Magnet system is superior to the CRY2/CIB1 system for application to gene expression systems.…”
Section: Figuresupporting
confidence: 87%
“…Screening methods and random mutations could help enhance photoswitch performance, as demonstrated by the iLID, TULIP, and FKF1 systems. Additionally, it should be possible to enhance the avidity of photoswitches by using assembled domains, such as CAD and ferritin, as demonstrated by the development of the CAD‐Magnet system by the Sato group …”
Section: Resultsmentioning
confidence: 99%
“…Light regulation also enables reversible protein activation/inactivation at the level of single cells or tissues . Photoswitches can also be activated within specific regions of interest: along the plasma membrane leading edge in an individual cell, for example, or in vivo within the liver . Conventional tools and methods such as inhibitors, RNAi, and constitutive active/dominant negative protein overexpression do not offer similar capabilities.…”
Section: Figurementioning
confidence: 99%
“…Cell membrane expansion was induced through PIP3 production upon blue light irradiation . The Sato group further improved the Magnet system by using the CAD system (CAD‐Magnet) . pMagFast2‐CAD was fused to the Tiam1 DH/PH domain, and CAAX was attached to nMagHigh1.…”
Section: Figurementioning
confidence: 99%
“…The activity of as ignaling compound can, for example, be directly perturbed to investigate the role of at arget protein in ac ellular functions uch as ruffling or lamellipodium/filopodiumf ormation.L ight regulation also enables reversiblep rotein activation/inactivation at the level of single cells [10] or tissues. [11] Photoswitches can also be activated within specific regions of interest:a long the plasma membrane leading edge in an individual cell, [10] for example, or in vivow ithin the liver. [12] Conventional tools and methodssuch as inhibitors, RNAi, and constitutive active/dominant negative protein overexpression do not offer similarc apabilities.…”
Signal transductions are the basis for all cellular functions. Previous studies investigating signal transductions mainly relied on pharmacological inhibition, RNA interference, and constitutive active/dominant negative protein expression systems. However, such studies do not allow the modulation of protein activity with high spatial and temporal precision in cells, tissues, and organs in animals. Recently, non-channelrhodopsin-type optogenetic tools for regulating signal transduction have emerged. These photoswitches address several disadvantages of previous techniques, and allow us to control a variety of signal transductions such as cell membrane dynamics, calcium signaling, lipid signaling, and apoptosis. In this review we summarize recent advances in the development of such photoswitches and in how these optotools are applied to signaling processes.
“…The Magnet system (pMag and nMag pair) derived from Vivid has the advantage of small size (151 aa), allowing rapid heterodimerization reversibility (half‐life is 6.8 s) . Additionally, a CAD‐Magnet system with increased avidity between pMag and nMag was developed and used to induce membrane ruffling . There are two reports that the Magnet system is superior to the CRY2/CIB1 system for application to gene expression systems.…”
Section: Figuresupporting
confidence: 87%
“…Screening methods and random mutations could help enhance photoswitch performance, as demonstrated by the iLID, TULIP, and FKF1 systems. Additionally, it should be possible to enhance the avidity of photoswitches by using assembled domains, such as CAD and ferritin, as demonstrated by the development of the CAD‐Magnet system by the Sato group …”
Section: Resultsmentioning
confidence: 99%
“…Light regulation also enables reversible protein activation/inactivation at the level of single cells or tissues . Photoswitches can also be activated within specific regions of interest: along the plasma membrane leading edge in an individual cell, for example, or in vivo within the liver . Conventional tools and methods such as inhibitors, RNAi, and constitutive active/dominant negative protein overexpression do not offer similar capabilities.…”
Section: Figurementioning
confidence: 99%
“…Cell membrane expansion was induced through PIP3 production upon blue light irradiation . The Sato group further improved the Magnet system by using the CAD system (CAD‐Magnet) . pMagFast2‐CAD was fused to the Tiam1 DH/PH domain, and CAAX was attached to nMagHigh1.…”
Section: Figurementioning
confidence: 99%
“…The activity of as ignaling compound can, for example, be directly perturbed to investigate the role of at arget protein in ac ellular functions uch as ruffling or lamellipodium/filopodiumf ormation.L ight regulation also enables reversiblep rotein activation/inactivation at the level of single cells [10] or tissues. [11] Photoswitches can also be activated within specific regions of interest:a long the plasma membrane leading edge in an individual cell, [10] for example, or in vivow ithin the liver. [12] Conventional tools and methodssuch as inhibitors, RNAi, and constitutive active/dominant negative protein overexpression do not offer similarc apabilities.…”
Signal transductions are the basis for all cellular functions. Previous studies investigating signal transductions mainly relied on pharmacological inhibition, RNA interference, and constitutive active/dominant negative protein expression systems. However, such studies do not allow the modulation of protein activity with high spatial and temporal precision in cells, tissues, and organs in animals. Recently, non-channelrhodopsin-type optogenetic tools for regulating signal transduction have emerged. These photoswitches address several disadvantages of previous techniques, and allow us to control a variety of signal transductions such as cell membrane dynamics, calcium signaling, lipid signaling, and apoptosis. In this review we summarize recent advances in the development of such photoswitches and in how these optotools are applied to signaling processes.
Optogenetics is a powerful method for studying dynamic processes in living cells and has advanced cell biology research over the recent past. Key to the successful application of optogenetics is the careful design of the light‐sensing module, typically employing a natural or engineered photoreceptor that links the exogenous light input to the cellular process under investigation. Light–oxygen–voltage (LOV) domains, a highly diverse class of small blue light sensors, have proven to be particularly versatile for engineering optogenetic input modules. These can function via diverse modalities, including inducible allostery, protein recruitment, dimerization, or dissociation. This study reviews recent advances in the development of LOV domain‐based optogenetic tools and their application for studying and controlling selected cellular functions. Focusing on the widely employed LOV2 domain from Avena sativa phototropin‐1, this review highlights the broad spectrum of engineering opportunities that can be explored to achieve customized optogenetic regulation. Finally, major bottlenecks in the development of optogenetic methods are discussed and strategies to overcome these with recent synthetic biology approaches are pointed out.
Since the neurobiological inception of optogenetics, light-controlled molecular perturbations have been applied in many scientific disciplines to both manipulate and observe cellular function. Proteins exhibiting light-sensitive conformational changes provide researchers with avenues for spatiotemporal control over the cellular environment and serve as valuable alternatives to chemically inducible systems. Optogenetic approaches have been developed to target proteins to specific subcellular compartments, allowing for the manipulation of nuclear translocation and plasma membrane morphology. Additionally, these tools have been harnessed for molecular interrogation of organelle function, location, and dynamics. Optogenetic approaches offer novel ways to answer fundamental biological questions and to improve the efficiency of bioengineered cell factories by controlling the assembly of synthetic organelles. This review first provides a summary of available optogenetic systems with an emphasis on their organelle-specific utility. It then explores the strategies employed for organelle targeting and concludes by discussing our perspective on the future of optogenetics to control subcellular structure and organization.
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