Droplet-based microfluidic analysis and screening of single plant cells

Yu, Ziyi; Boehm, Christian R.; Hibberd, Julian M.; Abell, Chris; Haseloff, Jim; Burgess, Steven J.; Reyna-Llorens, Ivan

doi:10.1101/199992

Cited by 3 publications

(3 citation statements)

References 43 publications

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“…Such approaches have mainly been applied in unicellular microorganisms. However, individual plant protoplasts can be captured within microfluidic chambers or spherical hydrogel beads (Nezhad, 2014; Grasso & Lintilhac, 2016) and microfluidic phenotyping of protoplasts has recently been reported (Yu et al , 2018). The development of on‐chip genetic manipulation and characterisation of plant cells is therefore technologically possible in the near‐term.…”

Section: Applying the Principles Of Engineering To Biologymentioning

confidence: 99%

Beyond natural: synthetic expansions of botanical form and function

Patron

2020

New Phytologist

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Summary Powered by developments that enabled genome‐scale investigations, systems biology emerged as a field aiming to understand how phenotypes emerge from network functions. These advances fuelled a new engineering discipline focussed on synthetic reconstructions of complex biological systems with the goal of predictable rational design and control. Initially, progress in the nascent field of synthetic biology was slow due to the ad hoc nature of molecular biology methods such as cloning. The application of engineering principles such as standardisation, together with several key technical advances, enabled a revolution in the speed and accuracy of genetic manipulation. Combined with mathematical and statistical modelling, this has improved the predictability of engineering biological systems of which nonlinearity and stochasticity are intrinsic features leading to remarkable achievements in biotechnology as well as novel insights into biological function. In the past decade, there has been slow but steady progress in establishing foundations for synthetic biology in plant systems. Recently, this has enabled model‐informed rational design to be successfully applied to the engineering of plant gene regulation and metabolism. Synthetic biology is now poised to transform the potential of plant biotechnology. However, reaching full potential will require conscious adjustments to the skillsets and mind sets of plant scientists.

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Section: Applying the Principles Of Engineering To Biologymentioning

confidence: 99%

Beyond natural: synthetic expansions of botanical form and function

Patron

2020

New Phytologist

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“…Microfluidics can compartmentalize single cells within monodisperse picolitre‐sized droplets in a cost‐effective and high‐throughput process, for example, screening of 5 × 10 7 individual reactions requires only 150 µl of reagents and 7 h at an estimated cost of only a few dollars, as demonstrated by Agresti et al (2010). Over the past decades, droplet microfluidics has enabled single‐cell analysis for a wide range of applications across biological science, biomedicine and biochemistry (Agresti et al, 2010; Brouzes et al, 2009; Yu et al, 2018). This is because (1) the extracellular environments are accurately mimicked (Hosokawa et al, 2017; Liu et al, 2020); (2) the genotype‐phenotype linkages are established at a single‐cell level (Bowman & Alper, 2019; Fischlechner et al, 2014; S. Li, Giardina, et al, 2018; M. Li, van Zee, Goda, et al, 2018); (3) the miniaturized confinement improves the detection limit (Agresti et al, 2010; Zhu et al, 2012); and (4) massive parallel analysis can be conducted to probe cellular heterogeneity (Headen et al, 2018; Hindson et al, 2011; Klein et al, 2015; Ostafe et al, 2014; Zinchenko et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

Microdroplet enabled cultivation of single yeast cells correlates with bulk growth and reveals subpopulation phenomena

Liu

Peng

et al. 2020

Biotech & Bioengineering

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Yeast has been engineered for cost‐effective organic acid production through metabolic engineering and synthetic biology techniques. However, cell growth assays in these processes were performed in bulk at the population level, thus obscuring the dynamics of rare single cells exhibiting beneficial traits. Here, we introduce the use of monodisperse picolitre droplets as bioreactors to cultivate yeast at the single‐cell level. We investigated the effect of acid stress on growth and the effect of potassium ions on propionic acid tolerance for single yeast cells of different species, genotypes, and phenotypes. The results showed that the average growth of single yeast cells in microdroplets experiences the same trend to those of yeast populations grown in bulk, and microdroplet compartments do not significantly affect cell viability. This approach offers the prospect of detecting cell‐to‐cell variations in growth and physiology and is expected to be applied for the engineering of yeast to produce value‐added bioproducts.

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“…Microfluidics can compartmentalize single cells within monodisperse picolitre-sized droplets in a cost-effective and high-throughput process, for example, screening of 5 × 10 7 individual reactions requires only 150 μL of reagents and seven hours at an estimated cost of only a few dollars, as demonstrated by Agresti et al (2010). Over the past decades, droplet microfluidics has enabled single-cell analysis for a wide range of applications across biological science, biomedicine and biochemistry (Agresti et al, 2010;Brouzes et al, 2009;Yu et al, 2018). This is because (1) the extracellular environments are accurately mimicked (Hosokawa et al, 2017;Liu et al, 2020); (2) the genotype-phenotype linkages are established at a single-cell level (Bowman and Alper, 2020;Fischlechner et al, 2014;Li et al, 2018aLi et al, , 2018b; (3) the miniaturized confinement improves the detection limit (Agresti et al, 2010;Zhu et al, 2012); and (4) massive parallel analysis can be conducted to probe cellular heterogeneity (Headen et al, 2018;Hindson et al, 2011;Klein et al, 2015;Ostafe et al, 2014;Zinchenko et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

Microdroplet enabled cultivation of single yeast cells correlates with bulk growth and reveals subpopulation phenomena

Liu

Peng

et al. 2020

Preprint

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Yeast has been engineered for cost-effective organic acid production through metabolic engineering and synthetic biology techniques. However, cell growth assays in these processes were performed in bulk at the population level, thus obscuring the dynamics of rare single cells exhibiting beneficial traits. Here, we introduce the use of monodisperse picolitre droplets as bioreactors to cultivate yeast at the single-cell level. We investigated the effect of acid stress on growth and the effect of potassium ions on propionic acid tolerance for single yeast cells of different species, genotypes and phenotypes. The results showed that the average growth of single yeast cells in microdroplets was identical to those of yeast populations grown in bulk, and microdroplet compartments do not significantly affect cell viability. This approach offers the prospect of detecting cell-to-cell variations in growth and physiology and is expected to be applied for the engineering of yeast to produce value-added bioproducts.

show abstract

Droplet-based microfluidic analysis and screening of single plant cells

Cited by 3 publications

References 43 publications

Beyond natural: synthetic expansions of botanical form and function

Beyond natural: synthetic expansions of botanical form and function

Microdroplet enabled cultivation of single yeast cells correlates with bulk growth and reveals subpopulation phenomena

Microdroplet enabled cultivation of single yeast cells correlates with bulk growth and reveals subpopulation phenomena

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