Fine tuning the LightOn light-switchable transgene expression system

Ma, Zhengcai; Du, Zengmin; Chen, Xianjun; Wang, Xue; Yang, Yi

doi:10.1016/j.bbrc.2013.09.092

Cited by 38 publications

(34 citation statements)

References 17 publications

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“…Notably, dynamic range of the effectors can be improved by tuning the thermodynamic properties of the LOV-Jα interaction 93 . Dimerization of VVD has been employed to light regulate the association of DNA binding domains and their recognition of target sequences (LightOn) 98,99 . Additionally, the LOV-containing FKF1 and GI (GIGANTEA) interaction domains have been joined to proteins of interest to allow light-triggered association of the fusions through the native FKF1-GI interaction 100 .…”

Section: Engineering Of Flavoprotein Light Sensorsmentioning

confidence: 99%

Photochemistry of flavoprotein light sensors

Conrad¹,

Manahan²,

Crane³

2014

Nat Chem Biol

205

234

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Three major classes of flavin photosensors, LOV domains, BLUF proteins and cryptochromes regulate diverse biological activities in response to blue-light. Recent studies of structure, spectroscopy and chemical mechanism have provided unprecedented insight into how each family operates at the molecular level. In general, the photoexcitation of the flavin cofactor leads to changes in redox and protonation states that ultimately remodel protein conformation and molecular interactions. For LOV domains, issues remain regarding early photochemical events, but common themes in conformational propagation have emerged across a diverse family of proteins. For BLUF proteins, photoinduced electron transfer reactions critical to light conversion are defined, but the subsequent rearrangement of hydrogen bonding networks key for signaling remain highly controversial. For cryptochromes, the relevant photocycles are actively debated, but mechanistic and functional studies are converging. Despite these challenges, our current understanding has enabled the engineering of flavoprotein photosensors for control of signaling processes within cells.

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Section: Engineering Of Flavoprotein Light Sensorsmentioning

confidence: 99%

Photochemistry of flavoprotein light sensors

Conrad¹,

Manahan²,

Crane³

2014

Nat Chem Biol

205

234

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“…For instance, when cumate was added in the medium, the elevated rtTAm level could enhance leaky expression of TRE3G promoter, in the absence of the other two inducers; when the cells were illuminated, increased mCherry-TCP interacted with leaky rtTAm, and resulted in higher rtTAm-dependent expression of luciferase. Previous study demonstrated the way to fine-tune basal and/or maximal expression of LightOn system [24], which provides insights for the future modification of the circuit, especially when stringent control of a specific promoter is needed. Similarly, lower leaky expression level would be achieved by modifying CMV5(CuO) promoter using another weaker enhancer element to replace the strong CMV enhancer or increase the level of CymR by using a stronger promoter.…”

Section: Discussionmentioning

confidence: 99%

Synthetic circuits that process multiple light and chemical signal inputs

Liu

Huang

2017

BMC Syst Biol

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BackgroundMulti-signal processing circuits are essential for rational design of sophisticated synthetic systems with good controllability and modularity, therefore, enable construction of high-level networks. Moreover, light-inducible systems provide fast and reversible means for spatiotemporal control of gene expression.ResultsHere, in HEK 293 cells, we present combinatory genetic circuits responding to light and chemical signals, simultaneously. We first constructed a dual input circuit converting different light intensities into varying of the sensitivity of the promoter to a chemical inducer (doxycycline). Next, we generated a ternary input circuit, which responded to light, doxycycline and cumate. This circuit allowed us to use different combinations of blue light and the two chemical inducers to generate gradual output values over two orders of magnitude.ConclusionsOverall, in this study, we devise genetic circuits sensing and processing light and chemical inducers. Our work may provide insights into bio-computation and fine-tuning expression of the transgene.Electronic supplementary materialThe online version of this article (doi:10.1186/s12918-016-0384-y) contains supplementary material, which is available to authorized users.

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“…We next examined whether M2 H37A can be used to ablate zebrafish neurons by establishing a system for Gal4/UAS-dependent expression in these cells. We generated heterozygous Tg(UAS:M2 H37A ;myl7:mCherry) zebrafish, using five tandem UAS sequences to minimize basal M2 H37A expression (Ma et al, 2013), and crossed them with a homozygous Tg(tuba1a:Gal4VP16;myl7:GFP) line. We then selected progeny that contained one copy of each transgene, which could be confirmed by the presence of both GFP and mCherry fluorescence in the heart (Fig.…”

Section: Genetically Encoded Neuron Ablation In Zebrafishmentioning

confidence: 99%

Targeted cell ablation in zebrafish using optogenetic transcriptional control

Mruk

Ciepla

Piza

et al. 2019

Preprint

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Cell ablation is a powerful method for elucidating the contributions of individual cell populations to embryonic development and tissue regeneration. Targeted cell loss in whole organisms has been typically achieved through expression of a cytotoxic or prodrug-activating gene product in the cell type of interest. This approach depends on the availability of tissue-specific promoters, and it does not allow further spatial selectivity within the promoter-defined region(s).To address this limitation, we have developed ablative methods that combine genetically encoded toxins, the tissue specificity afforded by cis-regulatory elements, and the conditionality of optogenetics. Using this integrative approach, we have ablated cells in zebrafish embryos with spatial and temporal precision.

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Fine tuning the LightOn light-switchable transgene expression system

Cited by 38 publications

References 17 publications

Photochemistry of flavoprotein light sensors

Photochemistry of flavoprotein light sensors

Synthetic circuits that process multiple light and chemical signal inputs

Targeted cell ablation in zebrafish using optogenetic transcriptional control

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