A modified single-cell electroporation method for molecule delivery into a motile protist, Euglena gracilis

Ohmachi, Masashi; Fujiwara, Yoshi; Muramatsu, Shuki; Yamada, Koji; Iwata, Osamu; Suzuki, Kengo; Wang, Dan Ohtan

doi:10.1016/j.mimet.2016.08.018

Cited by 10 publications

(6 citation statements)

References 20 publications

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“…This will be helpful for further basic studies on E. gracilis, which should include a long-lasting evaluation of a specific cell, e.g., observing its basic physiological phenomena, such as cell division ( Fig. S1 and Supplemental Movie), as well as the micro-manipulation of single cells, such as that through single-cell electroporation, to deliver genes and proteins to the cells (Ohmachi et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

Isolation and characterization of a motility-defective mutant ofEuglena gracilis

Muramatsu

Atsuji²,

Yamada³

et al. 2020

PeerJ

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Euglena gracilis is a green photosynthetic microalga that swims using its flagellum. This species has been used as a model organism for over half a century to study its metabolism and the mechanisms of its behavior. The development of mass-cultivation technology has led to E. gracilis application as a feedstock in various products such as foods. Therefore, breeding of E. gracilis has been attempted to improve the productivity of this feedstock for potential industrial applications. For this purpose, a characteristic that preserves the microalgal energy e.g., reduces motility, should be added to the cultivars. The objective of this study was to verify our hypothesis that E. gracilis locomotion-defective mutants are suitable for industrial applications because they save the energy required for locomotion. To test this hypothesis, we screened for E. gracilis mutants from Fe-ion-irradiated cell suspensions and established a mutant strain, ${\mathrm{M}}_{3}^{-}$ZFeL, which shows defects in flagellum formation and locomotion. The mutant strain exhibits a growth rate comparable to that of the wild type when cultured under autotrophic conditions, but had a slightly slower growth under heterotrophic conditions. It also stores 1.6 times the amount of paramylon, a crystal of β-1,3-glucan, under autotrophic culture conditions, and shows a faster sedimentation compared with that of the wild type, because of the deficiency in mobility and probably the high amount of paramylon accumulation. Such characteristics make E. gracilis mutant cells suitable for cost-effective mass cultivation and harvesting.

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Section: Discussionmentioning

confidence: 99%

Isolation and characterization of a motility-defective mutant ofEuglena gracilis

Muramatsu

Atsuji²,

Yamada³

et al. 2020

PeerJ

Self Cite

View full text Add to dashboard Cite

show abstract

“…In addition to the production cost, non-motile cells have the advantage of being easy to observe and manipulate in a limited area under the microscope. This will be helpful for further basic studies on E. gracilis, which should include a long-lasting evaluation of a specific cell, e.g., observing its basic physiological phenomena, such as cell division ( Figure S1 and Supplemental movie), as well as the micro-manipulation of single cells, such as that through single-cell electroporation, to deliver genes and proteins to the cells (Ohmachi et al 2016).…”

Section: Discussionmentioning

confidence: 99%

Peer Review #1 of "Isolation and characterization of a motility-defective mutant of Euglena gracilis (v0.1)"

González‐Halphen¹

2020

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Euglena gracilis is a green photosynthetic microalga that swims using its flagellum. This species has been used as a model organism for over half a century to study its metabolism and the mechanisms of its behavior. The development of mass-cultivation technology has led to E. gracilis application as a feedstock in various products such as foods. Therefore, breeding of E. gracilis has been attempted to improve the productivity of this feedstock for potential industrial applications. For this purpose, a characteristic that preserves the microalgal energy e.g., reduces motility, should be added to the cultivars. The objective of this study was to verify our hypothesis that E. gracilis locomotion-defective mutants are suitable for industrial applications because they save the energy required for locomotion. To test this hypothesis, we screened for E. gracilis mutants from Fe-ion-irradiated cell suspensions and established a mutant strain, M-3 ZFeL, which shows defects in flagellum formation and locomotion. The mutant strain exhibits a growth rate comparable to that of the wild type when cultured under autotrophic conditions, but had a slightly slower growth under heterotrophic conditions. It also stores 1.6 times the amount of paramylon, a crystal of β-1,3-glucan, under autotrophic culture conditions, and shows a faster sedimentation compared with that of the wild type, because of the deficiency in mobility and probably the high amount of paramylon accumulation. Such characteristics make E. gracilis mutant cells suitable for cost-effective mass cultivation and harvesting.

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“…A chloroplast transformation technique was developed in 2001 using biolistic transformation with confirmed transgene transcription (Doetsch et al, 2001). Later on, the focus was shifted to the nucleus, electroporation was developed and demonstrated to be a potential technique to transform fluorescent markers into the nucleus of E. gracilis (Ohmachi et al, 2016). Recently, a nuclear transformation technique with the help of Agrobacterium was also demonstrated to be successful in E. gracilis (Khatiwada et al, 2019;Becker et al, 2021).…”

Section: Potential Applications Of the Euglena Model In Metabolic Eng...mentioning

confidence: 99%

Metabolic network reconstruction of Euglena gracilis: Current state, challenges, and applications

Inwongwan

Pekkoh²,

Pumas³

et al. 2023

Front. Microbiol.

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A metabolic model, representing all biochemical reactions in a cell, is a prerequisite for several approaches in systems biology used to explore the metabolic phenotype of an organism. Despite the use of Euglena in diverse industrial applications and as a biological model, there is limited understanding of its metabolic network capacity. The unavailability of the completed genome data and the highly complex evolution of Euglena are significant obstacles to the reconstruction and analysis of its genome-scale metabolic model. In this mini-review, we discuss the current state and challenges of metabolic network reconstruction in Euglena gracilis. We have collated and present the available relevant data for the metabolic network reconstruction of E. gracilis, which could be used to improve the quality of the metabolic model of E. gracilis. Furthermore, we deliver the potential applications of the model in metabolic engineering. Altogether, it is supposed that this mini-review would facilitate the investigation of metabolic networks in Euglena and further lay out a direction for model-assisted metabolic engineering.

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A modified single-cell electroporation method for molecule delivery into a motile protist, Euglena gracilis

Cited by 10 publications

References 20 publications

Isolation and characterization of a motility-defective mutant ofEuglena gracilis

Isolation and characterization of a motility-defective mutant ofEuglena gracilis

Peer Review #1 of "Isolation and characterization of a motility-defective mutant of Euglena gracilis (v0.1)"

Metabolic network reconstruction of Euglena gracilis: Current state, challenges, and applications

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