Reconstruction of a chromatic response system in &lt;i&gt;Escherichia coli&lt;/i&gt;

Sugie, Yoshimi; Hori, Mayuko; Oka, Shunsuke; Ohtsuka, Hokuto; Aiba, Hirofumi

doi:10.2323/jgam.2016.01.006

Cited by 9 publications

(6 citation statements)

References 14 publications

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“…In previous work, we and others have engineered bacterial photoreceptors that regulate transcription in response to ultraviolet (maximal wavelength (λ max ) = 382–405 nm), blue (λ max = 450 nm), − green (λ max = 535 nm), − red (λ max = 650 nm), , and short wavelength near-infrared (NIR; λ max = 712 nm) light. Several of these photoreceptors have been used to spatially pattern gene expression across macroscopic ,, and microscopic cell populations, engineer a bacterial edge detector, induce permanent genetic memory via recombinase expression, program tailor-made expression dynamics of one , and two independent proteins, place protein expression under in silico feedback control, and reveal novel input/output dynamics of a widely used synthetic genetic circuit …”

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confidence: 99%

Engineering an E. coli Near-Infrared Light Sensor

2017

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Optogenetics is a technology wherein researchers combine light and genetically engineered photoreceptors to control biological processes with unrivaled precision. Near-infrared (NIR) wavelengths (>700 nm) are desirable optogenetic inputs due to their low phototoxicity and spectral isolation from most photoproteins. The bacteriophytochrome photoreceptor 1 (BphP1), found in several purple photosynthetic bacteria, senses NIR light and activates transcription of photosystem promoters by binding to and inhibiting the transcriptional repressor PpsR2. Here, we examine the response of a library of output promoters to increasing levels of Rhodopseudomonas palustris PpsR2 expression, and we identify that of Bradyrhizobium sp. BTAi1 crtE as the most strongly repressed in Escherichia coli. Next, we optimize Rps. palustris bphP1 and ppsR2 expression in a strain engineered to produce the required chromophore biliverdin IXα in order to demonstrate NIR-activated transcription. Unlike a previously engineered bacterial NIR photoreceptor, our system does not require production of a second messenger, and it exhibits rapid response dynamics. It is also the most red-shifted bacterial optogenetic tool yet reported by approximately 50 nm. Accordingly, our BphP1-PpsR2 system has numerous applications in bacterial optogenetics.

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confidence: 99%

Engineering an E. coli Near-Infrared Light Sensor

2017

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“…Though unforeseen, inactivity in response to blue light is not unprecedented for a CcaS protein. The CcaS homolog from Nostoc punctiforme also demonstrated blue-off behavior when repurposed as an optogenetic actuator in E. coli [44]. For CcaS HL , our studies suggest that this response potentially arises from the blue-light mediated activity of a second CcaS-associated pigment, a flavin.…”

Section: Discussionmentioning

confidence: 83%

Highlighter: an optogenetic actuator for light-mediated, high resolution gene expression control in plants

Larsen

Hofmann

Camacho

et al. 2022

Preprint

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Optogenetic actuators have revolutionized the resolution at which we can assert control over biological processes in living systems. In plants, deployment of optogenetics is challenging due to the need for these light-responsive systems to maintain a single activation state in conventional horticultural environments with light-dark cycling. Furthermore, many available optogenetic actuators are based on plant photoreceptors that might crosstalk with endogenous signaling processes, while others depend on exogenously supplied cofactors. To overcome such challenges, we have developed Highlighter; a synthetic, light-gated gene expression system tailored for in planta function. Highlighter is based on the photoswitchable CcaS-CcaR system from cyanobacteria and is repurposed for plants as a fully genetically encoded system, engineered to photoswitch with the endogenous plant chromophore, phytochromobilin. We deployed Highlighter in transiently transformed Nicotiana benthamiana for optogenetic control of fluorescent protein expression and innate immune responses. Using light to guide differential fluorescent protein expression in nuclei of neighboring cells, we demonstrate unprecedented spatiotemporal control of target gene expression. We furthermore regulate activation of plant immunity by modulating the spectral composition of white light, demonstrating optogenetic control of a biological process in horticultural light environments. Highlighter is a step forward for optogenetics in plants and a technology for high-resolution gene induction that will advance fundamental plant biology and provide new opportunities for crop improvement.

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“…The CcaS homolog from Nostoc punctiforme also demonstrated blue-off behavior when repurposed as an optogenetic actuator in E . coli [ 46 ] and for CcaS HL , our studies suggest that this response potentially arises from the blue-light-mediated activity of a second CcaS-associated pigment, a flavin.…”

Section: Discussionmentioning

confidence: 99%

Highlighter: An optogenetic system for high-resolution gene expression control in plants

Larsen,

Hofmann,

Camacho

et al. 2023

PLoS Biol

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Optogenetic actuators have revolutionized the resolution at which biological processes can be controlled. In plants, deployment of optogenetics is challenging due to the need for these light-responsive systems to function in the context of horticultural light environments. Furthermore, many available optogenetic actuators are based on plant photoreceptors that might crosstalk with endogenous signaling processes, while others depend on exogenously supplied cofactors. To overcome such challenges, we have developed Highlighter, a synthetic, light-gated gene expression system tailored for in planta function. Highlighter is based on the photoswitchable CcaS-CcaR system from cyanobacteria and is repurposed for plants as a fully genetically encoded system. Analysis of a re-engineered CcaS in Escherichia coli demonstrated green/red photoswitching with phytochromobilin, a chromophore endogenous to plants, but also revealed a blue light response likely derived from a flavin-binding LOV-like domain. We deployed Highlighter in transiently transformed Nicotiana benthamiana for optogenetic control of fluorescent protein expression. Using light to guide differential fluorescent protein expression in nuclei of neighboring cells, we demonstrate unprecedented spatiotemporal control of target gene expression. We implemented the system to demonstrate optogenetic control over plant immunity and pigment production through modulation of the spectral composition of broadband visible (white) light. Highlighter is a step forward for optogenetics in plants and a technology for high-resolution gene induction that will advance fundamental plant biology and provide new opportunities for crop improvement.

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Reconstruction of a chromatic response system in <i>Escherichia coli</i>

Cited by 9 publications

References 14 publications

Engineering an E. coli Near-Infrared Light Sensor

Engineering an E. coli Near-Infrared Light Sensor

Highlighter: an optogenetic actuator for light-mediated, high resolution gene expression control in plants

Highlighter: An optogenetic system for high-resolution gene expression control in plants

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