In vitro gene expression within membrane-free coacervate protocells

Tang, T-Y Dora; Swaay, Dirk van; deMello, Andrew J.; Anderson, J. L. Ross; Mann, Stephen

doi:10.1039/c5cc04220h

Cited by 164 publications

(61 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Such observations are in agreement with previous studies in bulk cell-free reactions where macromolecular crowding enhances transcription and impairs translation (Ge et al, 2011). To generate monodisperse coacervates in high throughput, Tang et al (2015) produced coacervates using a microfluidic device starting from a mixture of carboxymethyl-dextran/polylysine and TX-TL. However, they observed lower gene expression in coacervates compared to the bulk reaction, with results suggesting charge-induced precipitation of the reporter protein after its production.…”

Section: Liquid-liquid Phase Separationsupporting

confidence: 89%

Bottom-Up Construction of Complex Biomolecular Systems With Cell-Free Synthetic Biology

Laohakunakorn

Grasemann

Lavickova

et al. 2020

Front. Bioeng. Biotechnol.

104

101

View full text Add to dashboard Cite

Cell-free systems offer a promising approach to engineer biology since their open nature allows for well-controlled and characterized reaction conditions. In this review, we discuss the history and recent developments in engineering recombinant and crude extract systems, as well as breakthroughs in enabling technologies, that have facilitated increased throughput, compartmentalization, and spatial control of cell-free protein synthesis reactions. Combined with a deeper understanding of the cell-free systems themselves, these advances improve our ability to address a range of scientific questions. By mastering control of the cell-free platform, we will be in a position to construct increasingly complex biomolecular systems, and approach natural biological complexity in a bottom-up manner.

show abstract

Section: Liquid-liquid Phase Separationsupporting

confidence: 89%

Bottom-Up Construction of Complex Biomolecular Systems With Cell-Free Synthetic Biology

Laohakunakorn

Grasemann

Lavickova

et al. 2020

Front. Bioeng. Biotechnol.

104

101

View full text Add to dashboard Cite

show abstract

“…The formation of a coacervate phase has been demonstrated with a variety of polyionic molecules, most notably with use of cationic peptides or unnatural polymers in combination with nucleotides, RNA, or fatty acids, or with use of elastin‐like polypeptides . In addition, complex coacervates can readily be loaded with a wide range of cargos, including proteins, nucleic acids, and metabolites, and have been shown to support various cell‐like processes such as RNA folding and ribosomal activity, transcription and translation, and dissipative self‐assembly of cytoskeletal proteins . This makes complex coacervates ideal mimics both for the liquid organelles that perform specialized functions in the cell, such as nucleoli, and for the cell cytosol…”

Section: Introductionmentioning

confidence: 99%

Physicochemical Characterization of Polymer‐Stabilized Coacervate Protocells

et al. 2019

View full text Add to dashboard Cite

The bottom‐up construction of cell mimics has produced a range of membrane‐bound protocells that have been endowed with functionality and biochemical processes reminiscent of living systems. The contents of these compartments, however, experience semidilute conditions, whereas macromolecules in the cytosol exist in protein‐rich, crowded environments that affect their physicochemical properties, such as diffusion and catalytic activity. Recently, complex coacervates have emerged as attractive protocellular models because their condensed interiors would be expected to mimic this crowding better. Here we explore some relevant physicochemical properties of a recently developed polymer‐stabilized coacervate system, such as the diffusion of macromolecules in the condensed coacervate phase, relative to in dilute solutions, the buffering capacity of the core, the molecular organization of the polymer membrane, the permeability characteristics of this membrane towards a wide range of compounds, and the behavior of a simple enzymatic reaction. In addition, either the coacervate charge or the cargo charge is engineered to allow the selective loading of protein cargo into the coacervate protocells. Our in‐depth characterization has revealed that these polymer‐stabilized coacervate protocells have many desirable properties, thus making them attractive candidates for the investigation of biochemical processes in stable, controlled, tunable, and increasingly cell‐like environments.

show abstract

“…Compartments can even be made without a membrane at all [20]. Neither aqueous two-phase systems [21,22], polyelectrolyte polymer containing coacervates [23], hydrogels [24], microfluidic chip-based compartments [25,26], nor bead-based systems [27] contain a membrane.…”

Section: Chemical Communication Between Artificial Cellsmentioning

confidence: 99%

Toward long-lasting artificial cells that better mimic natural living cells

Martín

Valer

Mansy

2019

Emerging Topics in Life Sciences

View full text Add to dashboard Cite

Chemical communication is ubiquitous in biology, and so efforts in building convincing cellular mimics must consider how cells behave on a population level. Simple model systems have been built in the laboratory that show communication between different artificial cells and artificial cells with natural, living cells. Examples include artificial cells that depend on purely abiological components and artificial cells built from biological components and are driven by biological mechanisms. However, an artificial cell solely built to communicate chemically without carrying the machinery needed for self-preservation cannot remain active for long periods of time. What is needed is to begin integrating the pathways required for chemical communication with metabolic-like chemistry so that robust artificial systems can be built that better inform biology and aid in the generation of new technologies.

show abstract

In vitro gene expression within membrane-free coacervate protocells

Abstract: Cell-free gene expression of a fluorescent protein (mCherry) is demonstrated within the molecularly crowded matrix of a polysaccharide/polypeptide coacervate.

Cited by 164 publications

References 29 publications

Bottom-Up Construction of Complex Biomolecular Systems With Cell-Free Synthetic Biology

Bottom-Up Construction of Complex Biomolecular Systems With Cell-Free Synthetic Biology

Physicochemical Characterization of Polymer‐Stabilized Coacervate Protocells

Toward long-lasting artificial cells that better mimic natural living cells

Contact Info

Product

Resources

About