Optogenetics for transcriptional programming and genetic engineering

Lan, Tian; He, Lian; Huang, Yun; Zhou, Yubin

doi:10.1016/j.tig.2022.05.014

Cited by 30 publications

(28 citation statements)

References 127 publications

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“…Optogenetics is a powerful technique that allows for dynamic, spatial, and temporal control over cellular behavior using light. − Optogenetics leverages light-sensitive proteins, taking advantage of light responsive changes in protein conformation to actuate processes inside the cell. , Such tools have been used to activate specific signaling pathways, − repress and activate transcription, − control protein localization, − and induce protein degradation. , A common approach is to control a process of interest by fusing effectors to light-activated hetero- or homodimerizers to generate activity through proximity. For example, light-sensitive split transcription factors (TFs) are frequently generated by fusing one protein of an optical heterodimerizer pair to a DNA-binding domain (DBD) and the other to an activation domain (AD).…”

Section: Introductionmentioning

confidence: 99%

Lustro: High-Throughput Optogenetic Experiments Enabled by Automation and a Yeast Optogenetic Toolkit

Harmer

McClean

2023

ACS Synth. Biol.

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Optogenetic systems use genetically encoded lightsensitive proteins to control cellular processes. This provides the potential to orthogonally control cells with light; however, these systems require many design−build−test cycles to achieve a functional design and multiple illumination variables need to be laboriously tuned for optimal stimulation. We combine laboratory automation and a modular cloning scheme to enable highthroughput construction and characterization of optogenetic split transcription factors in Saccharomyces cerevisiae. We expand the yeast optogenetic toolkit to include variants of the cryptochromes and enhanced Magnets, incorporate these light-sensitive dimerizers into split transcription factors, and automate illumination and measurement of cultures in a 96-well microplate format for high-throughput characterization. We use this approach to rationally design and test an optimized enhanced Magnet transcription factor with improved light-sensitive gene expression. This approach is generalizable to the high-throughput characterization of optogenetic systems across a range of biological systems and applications.

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Section: Introductionmentioning

confidence: 99%

Lustro: High-Throughput Optogenetic Experiments Enabled by Automation and a Yeast Optogenetic Toolkit

Harmer

McClean

2023

ACS Synth. Biol.

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“…Photoswitchable probes have been developing rapidly in neuroscience, immunity, and other biological science fields over the past decade, especially since the algal protein channelrhodopsin-2 was first found to be expressed in mammalian neurons in response to light by Edward S. Boyden [ 46 , 47 , 48 ]. These tools show their power in regulating signaling networks with many advantages such as a superior temporal and spatial resolution, easy delivery, rapid reversibility, and fewer off-target side effects [ 49 , 50 ].…”

Section: Introductionmentioning

confidence: 99%

Coelenterazine-Type Bioluminescence-Induced Optical Probes for Sensing and Controlling Biological Processes

Jiang

Song

Zhang

2023

IJMS

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Bioluminescence-based probes have long been used to quantify and visualize biological processes in vitro and in vivo. Over the past years, we have witnessed the trend of bioluminescence-driven optogenetic systems. Typically, bioluminescence emitted from coelenterazine-type luciferin–luciferase reactions activate light-sensitive proteins, which induce downstream events. The development of coelenterazine-type bioluminescence-induced photosensory domain-based probes has been applied in the imaging, sensing, and control of cellular activities, signaling pathways, and synthetic genetic circuits in vitro and in vivo. This strategy can not only shed light on the mechanisms of diseases, but also promote interrelated therapy development. Here, this review provides an overview of these optical probes for sensing and controlling biological processes, highlights their applications and optimizations, and discusses the possible future directions.

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“…Optogenetics is a powerful technique that allows for dynamic, spatial, and temporal control over cellular behavior using light [1][2][3] . Optogenetics leverages light-sensitive proteins, taking advantage of light responsive changes in protein conformation to actuate processes inside the cell 4,5 .…”

Section: Introductionmentioning

confidence: 99%

Lustro: High-throughput optogenetic experiments enabled by automation and a yeast optogenetic toolkit

Harmer

McClean

2023

Preprint

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Optogenetic systems use genetically-encoded light-sensitive proteins to control cellular processes. This provides the potential to orthogonally control cells with light, however these systems require many design-build-test cycles to achieve a functional design and multiple illumination variables need to be laboriously tuned for optimal stimulation. We combine laboratory automation and a modular cloning scheme to enable high-throughput construction and characterization of optogenetic split transcription factors in Saccharomyces cerevisiae. We expand the yeast optogenetic toolkit to include variants of the cryptochromes and Enhanced Magnets, incorporate these light-sensitive dimerizers into split transcription factors, and automate illumination and measurement of cultures in a 96-well microplate format for high-throughput characterization. We use this approach to rationally design and test an optimized Enhanced Magnet transcription factor with improved light-sensitive gene expression. This approach is generalizable to high-throughput characterization of optogenetic systems across a range of biological systems and applications.

show abstract

Optogenetics for transcriptional programming and genetic engineering

Cited by 30 publications

References 127 publications

Lustro: High-Throughput Optogenetic Experiments Enabled by Automation and a Yeast Optogenetic Toolkit

Lustro: High-Throughput Optogenetic Experiments Enabled by Automation and a Yeast Optogenetic Toolkit

Coelenterazine-Type Bioluminescence-Induced Optical Probes for Sensing and Controlling Biological Processes

Lustro: High-throughput optogenetic experiments enabled by automation and a yeast optogenetic toolkit

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