Optimized light-inducible transcription in mammalian cells using Flavin Kelch-repeat F-box1/GIGANTEA and CRY2/CIB1

Quejada, Jose R.; Park, Seon-Hye E.; Awari, Daniel Wasim; Shi, Fan; Yamamoto, Hannah E.; Kawano, Fuun; Jung, Juergen C.; Yazawa, Masayuki

doi:10.1093/nar/gkx804

Cited by 32 publications

(34 citation statements)

References 29 publications

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“…[9c] Recently,t he Yazawa group generated an ew FKF1 mutant (H105L) by mutagenesis. [27] Blue-light-induced dimerization between FKF1(H105L) and GI resulted in enhanced gene expression with the Gal4/VP16 system,r elative to ap revious combination using FKF1(G128D) and GI. However,ad rawback of the FKF1/GIs ystem is that these are large proteins (619 and 1173 amino acids, respectively) in comparison with other bluelight-induced dimerization systems, including TULIPsa nd iLIDs, and this could decrease expression levels and thus lessen the switching power of this dimerizationsystem.…”

Section: Fkf1/giganteamentioning

confidence: 99%

“…The Dolmetsch group harnessed this dimerization property to develop a technology for gene expression systems (Figure G) and for translocating proteins from the cytosol to the membrane in a living cell . Recently, the Yazawa group generated a new FKF1 mutant (H105L) by mutagenesis . Blue‐light‐induced dimerization between FKF1(H105L) and GI resulted in enhanced gene expression with the Gal4/VP16 system, relative to a previous combination using FKF1(G128D) and GI.…”

Section: Figurementioning

confidence: 99%

“…). Leakiness in CRY2/CIB1 has been reported by demonstration that CRY2/CIB1 exhibits higher basal signal intensity under dark conditions than the FKF1/GI system for application to gene expression systems . Additionally, CRY2PHR induces unwanted gene expression in the dark state unlike full‐length CRY2 .…”

Section: Figurementioning

confidence: 99%

“…Leakiness in CRY2/CIB1 has been reported by demonstration that CRY2/CIB1 exhibits higherb asal signal intensityu nder dark conditions than the FKF1/GIs ystem fora pplication to gene expression systems. [27] Additionally,C RY2PHR induces unwanted gene expressioni nt he dark state unlike full-length CRY2. [30] Thus, when some leakiness of signal is acceptable, the CRY2/CIB1 system is appropriate as af irst-choice photoswitch.…”

Section: Comparison Of the Photoswitchesmentioning

confidence: 99%

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Induction of Signal Transduction by Using Non‐Channelrhodopsin‐Type Optogenetic Tools

Ueda

Sato

2018

ChemBioChem

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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.

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Section: Fkf1/giganteamentioning

confidence: 99%

Section: Figurementioning

confidence: 99%

Section: Figurementioning

confidence: 99%

Section: Comparison Of the Photoswitchesmentioning

confidence: 99%

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Induction of Signal Transduction by Using Non‐Channelrhodopsin‐Type Optogenetic Tools

Ueda

Sato

2018

ChemBioChem

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“…For instance, channelrhodopsin 1 is a light-gated ion channel that responds to blue light to allow for experimental control over neuronal firing. Similarly, cryptochrome 2 (Cry2) [3][4][5] and light-oxygen sensitive protein (LOV) based systems [6][7][8] utilize blue light to regulate protein binding and gene expression. Additionally, geneticallyencoded calcium sensor technologies to visualize neuronal activity states are becoming more widely utilized both in vivo and in vitro, and these sensors often rely on prolonged or repeated blue light exposure [9][10][11] .…”

mentioning

confidence: 99%

Blue light-induced gene expression alterations in cultured neurons are the result of phototoxic interactions with neuronal culture media

Duke

Savell

Phillips

et al. 2019

Preprint

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Blue waveform light is used as an optical actuator in numerous optogenetic technologies employed in neuronal systems. However, the potential side effects of blue waveform light in neurons has not been thoroughly explored, and recent reports suggest that neuronal exposure to blue light can induce transcriptional alterations in vitro and in vivo. Here, we examined the effects of blue waveform light in cultured primary rat cortical neurons. Exposure to blue light (470nm) resulted in upregulation of several immediate early genes (IEGs) traditionally used as markers of neuronal activity, including Fos and Fosb, but did not alter the expression of circadian clock genes Bmal1, Cry1, Cry2, Clock, or Per2. IEG expression was increased following 4 hours of 5% duty cycle light exposure, and IEG induction was not dependent on light pulse width. Elevated levels of blue light exposure induced a loss of cell viability in vitro, suggestive of overt phototoxicity. Changes in gene expression induced by blue waveform light were prevented when neurons were cultured in a photoinert media supplemented with a photostable neuronal supplement instead of commonly utilized neuronal culture media and supplements. Together, these findings suggest that light-induced gene expression alterations observed in vitro stem from a phototoxic interaction between commonly used media and neurons, and offer a solution to prevent this toxicity when using photoactivatable technology in vitro.

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Synthetic Gene Circuits for Regulation of Next‐Generation Cell‐Based Therapeutics

Teixeira,

Fussenegger

2023

Advanced Science

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Arming human cells with synthetic gene circuits enables to expand their capacity to execute superior sensing and response actions, offering tremendous potential for innovative cellular therapeutics. This can be achieved by assembling components from an ever‐expanding molecular toolkit, incorporating switches based on transcriptional, translational, or post‐translational control mechanisms. This review provides examples from the three classes of switches, and discusses their advantages and limitations to regulate the activity of therapeutic cells in vivo. Genetic switches designed to recognize internal disease‐associated signals often encode intricate actuation programs that orchestrate a reduction in the sensed signal, establishing a closed‐loop architecture. Conversely, switches engineered to detect external molecular or physical cues operate in an open‐loop fashion, switching on or off upon signal exposure. The integration of such synthetic gene circuits into the next generation of chimeric antigen receptor T‐cells is already enabling precise calibration of immune responses in terms of magnitude and timing, thereby improving the potency and safety of therapeutic cells. Furthermore, pre‐clinical engineered cells targeting other chronic diseases are gathering increasing attention, and this review discusses the path forward for achieving clinical success. With synthetic biology at the forefront, cellular therapeutics holds great promise for groundbreaking treatments.

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Optimized light-inducible transcription in mammalian cells using Flavin Kelch-repeat F-box1/GIGANTEA and CRY2/CIB1

Cited by 32 publications

References 29 publications

Induction of Signal Transduction by Using Non‐Channelrhodopsin‐Type Optogenetic Tools

Induction of Signal Transduction by Using Non‐Channelrhodopsin‐Type Optogenetic Tools

Blue light-induced gene expression alterations in cultured neurons are the result of phototoxic interactions with neuronal culture media

Synthetic Gene Circuits for Regulation of Next‐Generation Cell‐Based Therapeutics

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