Light-mediated control of Gene expression in mammalian cells

Yamada, Mayumi; Nagasaki, Shinji C.; Ozawa, Toshio; Imayoshi, Itaru

doi:10.1016/j.neures.2019.12.018

Cited by 33 publications

(25 citation statements)

References 115 publications

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“…Thus, these systems are not applicable for controlling the dynamics of regulators in most pulsing systems. To overcome these limitations, a series of light-inducible systems have been developed that allow control of gene expression with unprecedented spatial and temporal precision 12,13 . These optogenetic systems provide robust and convenient tools to achieve pulsatile dynamic regulation of regulators, allowing direct comparison between functional outcomes and concentration-time relationships of external stimuli.…”

mentioning

confidence: 99%

A synthetic BRET-based optogenetic device for pulsatile transgene expression enabling glucose homeostasis in mice

Chen

Qian

et al. 2021

Nat Commun

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Pulsing cellular dynamics in genetic circuits have been shown to provide critical capabilities to cells in stress response, signaling and development. Despite the fascinating discoveries made in the past few years, the mechanisms and functional capabilities of most pulsing systems remain unclear, and one of the critical challenges is the lack of a technology that allows pulsatile regulation of transgene expression both in vitro and in vivo. Here, we describe the development of a synthetic BRET-based transgene expression (LuminON) system based on a luminescent transcription factor, termed luminGAVPO, by fusing NanoLuc luciferase to the light-switchable transcription factor GAVPO. luminGAVPO allows pulsatile and quantitative activation of transgene expression via both chemogenetic and optogenetic approaches in mammalian cells and mice. Both the pulse amplitude and duration of transgene expression are highly tunable via adjustment of the amount of furimazine. We further demonstrated LuminON-mediated blood-glucose homeostasis in type 1 diabetic mice. We believe that the BRET-based LuminON system with the pulsatile dynamics of transgene expression provides a highly sensitive tool for precise manipulation in biological systems that has strong potential for application in diverse basic biological studies and gene- and cell-based precision therapies in the future.

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mentioning

confidence: 99%

A synthetic BRET-based optogenetic device for pulsatile transgene expression enabling glucose homeostasis in mice

Chen

Qian

et al. 2021

Nat Commun

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“…For example, the dual-controlled “Blue-OFF” system combines a KRAB-EL222 light-inducible repressor with a B-LID (blue light-inducible degradation domain) protein degradation module, to rapidly downregulate target protein levels in response to blue light ( Baaske et al, 2018 ). By combining such systems with recently developed light-inducible gene expression tools ( Yamada et al, 2020 ), blue light could be used to trigger both cell-cycle regulator degradation (to halt proliferation) and expression of the product gene (to start the production phase).…”

Section: Novel Synthetic Biology Methods For Proliferation Controlmentioning

confidence: 99%

Decoupling Growth and Protein Production in CHO Cells: A Targeted Approach

Donaldson

Dale

Rosser

2021

Front. Bioeng. Biotechnol.

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Fed-batch cultures of Chinese Hamster Ovary cells have been used to produce high quantities of biotherapeutics, particularly monoclonal antibodies. However, a growing number of next-generation biotherapeutics, such as bi-specific antibodies and fusion proteins, are difficult to express using standard fed-batch processes. Decoupling cell growth and biotherapeutic production is becoming an increasingly desired strategy for the biomanufacturing industry, especially for difficult-to-express products. Cells are grown to a high cell density in the absence of recombinant protein production (the growth phase), then expression of the recombinant protein is induced and cell proliferation halted (the production phase), usually by combining an inducible gene expression system with a proliferation control strategy. Separating the growth and production phases allows cell resources to be more efficiently directed toward either growth or production, improving growth characteristics and enhancing the production of difficult to express proteins. However, current mammalian cell proliferation control methods rely on temperature shifts and chemical agents, which interact with many non-proliferation pathways, leading to variable impacts on product quality and culture viability. Synthetic biology offers an alternative approach by strategically targeting proliferation pathways to arrest cell growth but have largely remained unused in industrial bioproduction. Due to recent developments in microbial decoupling systems and advances in available mammalian cell engineering tools, we propose that the synthetic biology approach to decoupling growth and production needs revisiting.

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“…However, neither were affected by the increased interneuron activity, thus supporting previous findings indicating that postnatal GABAergic activity does not affect cortical oligodendroglia (Balia et al, 2017 ; Ortolani et al, 2018 ). Optogenetics also allows the control of gene expression via light exposure (Yamada et al, 2020 ) and modulation of gene expression in OPCs in vivo by shining light in a region-specific manner is an approach that could be used in the future.…”

Section: Functional Physiological Approachesmentioning

confidence: 99%

Novel Tools and Investigative Approaches for the Study of Oligodendrocyte Precursor Cells (NG2-Glia) in CNS Development and Disease

Galichet

Clayton

Lovell‐Badge

2021

Front. Cell. Neurosci.

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Oligodendrocyte progenitor cells (OPCs), also referred to as NG2-glia, are the most proliferative cell type in the adult central nervous system. While the primary role of OPCs is to serve as progenitors for oligodendrocytes, in recent years, it has become increasingly clear that OPCs fulfil a number of other functions. Indeed, independent of their role as stem cells, it is evident that OPCs can regulate the metabolic environment, directly interact with and modulate neuronal function, maintain the blood brain barrier (BBB) and regulate inflammation. In this review article, we discuss the state-of-the-art tools and investigative approaches being used to characterize the biology and function of OPCs. From functional genetic investigation to single cell sequencing and from lineage tracing to functional imaging, we discuss the important discoveries uncovered by these techniques, such as functional and spatial OPC heterogeneity, novel OPC marker genes, the interaction of OPCs with other cells types, and how OPCs integrate and respond to signals from neighboring cells. Finally, we review the use of in vitro assay to assess OPC functions. These methodologies promise to lead to ever greater understanding of this enigmatic cell type, which in turn will shed light on the pathogenesis and potential treatment strategies for a number of diseases, such as multiple sclerosis (MS) and gliomas.

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Light-mediated control of Gene expression in mammalian cells

Cited by 33 publications

References 115 publications

A synthetic BRET-based optogenetic device for pulsatile transgene expression enabling glucose homeostasis in mice

A synthetic BRET-based optogenetic device for pulsatile transgene expression enabling glucose homeostasis in mice

Decoupling Growth and Protein Production in CHO Cells: A Targeted Approach

Novel Tools and Investigative Approaches for the Study of Oligodendrocyte Precursor Cells (NG2-Glia) in CNS Development and Disease

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