Optogenetic strategies for the control of gene expression in yeasts

Pérez, Ana Laura A.; Piva, Luiza Cesca; Fulber, Julia P.C.; Moraes, Lídia Maria Pepe de; Marco, Janice Lisboa De; Vieira, Hugo L.A.; Coelho, Cíntia Marques; Reis, Viviane Castelo Branco; Torres, Fernando Araripe Gonçalves

doi:10.1016/j.biotechadv.2021.107839

Cited by 19 publications

(14 citation statements)

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“…A plethora of alternative promoters, both inducible and constitutive, have been developed for all the yeasts discussed here. Promoters induced by light are beginning to be tested in S. cerevisiae , and are outperforming traditional ones (i.e., carbon source‐induced promoters) (Pérez et al., 2022; Salinas et al., 2018). For Kl.…”

Section: Expression Hostsmentioning

confidence: 99%

Precision cellular agriculture: The future role of recombinantly expressed protein as food

Dupuis

Cheung

Newman

et al. 2022

Comp Rev Food Sci Food Safe

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Cellular agriculture is a rapidly emerging field, within which cultured meat has attracted the majority of media attention in recent years. An equally promising area of cellular agriculture, and one that has produced far more actual food ingredients that have been incorporated into commercially available products, is the use of cellular hosts to produce soluble proteins, herein referred to as precision cellular agriculture (PCAg). In PCAg, specific animal‐ or plant‐sourced proteins are expressed recombinantly in unicellular hosts—the majority of which are yeast—and harvested for food use. The numerous advantages of PCAg over traditional agriculture, including a smaller carbon footprint and more consistent products, have led to extensive research on its utility. This review is the first to survey proteins currently being expressed using PCAg for food purposes. A growing number of viable expression hosts and recent advances for increased protein yields and process optimization have led to its application for producing milk, egg, and muscle proteins; plant hemoglobin; sweet‐tasting plant proteins; and ice‐binding proteins. Current knowledge gaps present research opportunities for optimizing expression hosts, tailoring posttranslational modifications, and expanding the scope of proteins produced. Considerations for the expansion of PCAg and its implications on food regulation, society, ethics, and the environment are also discussed. Considering the current trajectory of PCAg, food proteins from any biological source can likely be expressed recombinantly and used as purified food ingredients to create novel and tailored food products.

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Section: Expression Hostsmentioning

confidence: 99%

Precision cellular agriculture: The future role of recombinantly expressed protein as food

Dupuis

Cheung

Newman

et al. 2022

Comp Rev Food Sci Food Safe

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“…Recent progress in synthetic biology has established light as a leading tool for observation as well as stimulation of synthetic constructs in cellular contexts ( MacDonald and Deans, 2016 ; Toettcher et al, 2011a ; Deisseroth, 2011 ; Müller et al, 2015 ; Krueger et al, 2019 ; Baumschlager and Khammash, 2021 ; Gheorghiu et al, 2021 ; Forlani and Di Ventura, 2021 ; Pérez et al, 2021 ). Light can be precisely localized in space and instantaneously turned on and off.…”

Section: Introductionmentioning

confidence: 99%

“…This use of light to influence cellular processes is commonly referred to as optogenetics ( Toettcher et al, 2011a ; Deisseroth, 2011 ). Several optogenetic tools, pertaining to different cell types and spanning different wavelengths of light, have been developed and studied in the last decade ( Tischer and Weiner, 2014 ; Müller et al, 2015 ; Repina et al, 2017 ; Kolar et al, 2018 ; Baumschlager and Khammash, 2021 ; Gheorghiu et al, 2021 ; Forlani and Di Ventura, 2021 ; Pérez et al, 2021 ). Applications of these optogenetic tools have been expanding rapidly in various disciplines, such as developmental biology ( Johnson and Toettcher, 2018 ; Krueger et al, 2019 ).…”

Section: Introductionmentioning

confidence: 99%

Platforms for Optogenetic Stimulation and Feedback Control

Kumar

Khammash²

2022

Front. Bioeng. Biotechnol.

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Harnessing the potential of optogenetics in biology requires methodologies from different disciplines ranging from biology, to mechatronics engineering, to control engineering. Light stimulation of a synthetic optogenetic construct in a given biological species can only be achieved via a suitable light stimulation platform. Emerging optogenetic applications entail a consistent, reproducible, and regulated delivery of light adapted to the application requirement. In this review, we explore the evolution of light-induction hardware-software platforms from simple illumination set-ups to sophisticated microscopy, microtiter plate and bioreactor designs, and discuss their respective advantages and disadvantages. Here, we examine design approaches followed in performing optogenetic experiments spanning different cell types and culture volumes, with induction capabilities ranging from single cell stimulation to entire cell culture illumination. The development of automated measurement and stimulation schemes on these platforms has enabled researchers to implement various in silico feedback control strategies to achieve computer-controlled living systems—a theme we briefly discuss in the last part of this review.

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“…First, except for the Phr1 photolyase, the genome of S. cerevisiae lacks functional photoreceptors . Thus, plenty of research has reported the successful implementation of optogenetic switches in yeast, where light seems to act as an orthogonal input. , On the other hand, the life cycle of S. cerevisiae involves a communication mechanism based on small diffusible pheromones, which serve as signaling molecules. , The binding of these pheromones to specific G protein-coupled receptors in the cell surface triggers the mating process, which involves the activation of a mitogen-activated protein kinase (MAPK) cascade leading to chemotropic growth by large-scale changes in gene expression, cell division cycle, and morphology. − As S. cerevisiae lacks motility mechanisms, this pheromone-dependent polarized extension allows haploid yeast cells to get close, make contact, and fuse to form a diploid zygote. , …”

Section: Introductionmentioning

confidence: 99%

Coupling Cell Communication and Optogenetics: Implementation of a Light-Inducible Intercellular System in Yeast

Rojas

Larrondo

2022

ACS Synth. Biol.

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Cell communication is a widespread mechanism in biology, allowing the transmission of information about environmental conditions. In order to understand how cell communication modulates relevant biological processes such as survival, division, differentiation, and apoptosis, different synthetic systems based on chemical induction have been successfully developed. In this work, we coupled cell communication and optogenetics in the budding yeast Saccharomyces cerevisiae. Our approach is based on two strains connected by the light-dependent production of α-factor pheromone in one cell type, which induces gene expression in the other type. After the individual characterization of the different variants of both strains, the optogenetic intercellular system was evaluated by combining the cells under contrasting illumination conditions. Using luciferase as a reporter gene, specific co-cultures at a 1:1 ratio displayed activation of the response upon constant blue light, which was not observed for the same cell mixtures grown in darkness. Then, the system was assessed at several dark/blue-light transitions, where the response level varies depending on the moment in which illumination was delivered. Furthermore, we observed that the amplitude of response can be tuned by modifying the initial ratio between both strains. Finally, the two-population system showed higher fold inductions in comparison with autonomous strains. Altogether, these results demonstrated that external light information is propagated through a diffusible signaling molecule to modulate gene expression in a synthetic system involving microbial cells, which will pave the road for studies allowing optogenetic control of population-level dynamics.

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Optogenetic strategies for the control of gene expression in yeasts

Cited by 19 publications

References 100 publications

Precision cellular agriculture: The future role of recombinantly expressed protein as food

Precision cellular agriculture: The future role of recombinantly expressed protein as food

Platforms for Optogenetic Stimulation and Feedback Control

Coupling Cell Communication and Optogenetics: Implementation of a Light-Inducible Intercellular System in Yeast

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