Coacervates have been widely studied as model compartments in protocell research. Complex coacervates composed of disordered proteins and RNA have also been shown to play an important role in ...cellular processes. Herein, we report on a microfluidic strategy for constructing monodisperse coacervate droplets encapsulated within uniform unilamellar liposomes. These structures represent a bottom‐up approach to hierarchically structured protocells, as demonstrated by storage and release of DNA from the encapsulated coacervates as well as localized transcription.
Artificial organelles: Monodisperse liposomes containing monodisperse dynamic, functional compartments were achieved using microfluidics and employed as artificial/protocell models to imitate membrane‐less intracellular organelle‐based compartmentalization.
Complex coacervates are water droplets dispersed in water, which are formed by spontaneous liquid–liquid phase separation of an aqueous solution of two oppositely charged polyelectrolytes. Similar to ...the membraneless organelles that exist in biological cells, complex coacervate droplets are membraneless and have a myriad of features including easy formation, high viscosity, selective encapsulation of biomolecules, and dynamic behaviors in response to environmental stimuli, which make coacervates an excellent option for constructing artificial membraneless organelles. In this article, I first summarize recent advances in artificial compartments that are built from coacervates and their response to changes in the surrounding environment and then show the advantages of microfluidic techniques in the preparation of monodisperse coacervates and encapsulation of coacervates in droplets and liposomes to construct complex cell-like compartments, and finally discuss the future challenges of such membraneless aqueous compartments in cell mimics and origin of life.
Vesosomes are nested liposomal structures with high potential as advanced drug delivery vehicles, bioreactors and artificial cells. However, to date no method has been reported to prepare ...monodisperse vesosomes of controlled size. Here we report on a multistep microfluidic strategy for hierarchically assembling uniform vesosomes from dewetting of double emulsion templates. The control afforded by our method is illustrated by the formation of concentric, pericentric and multicompartment liposomes. The microfluidic route to vesosomes offers an exceptional platform to build artificial cells as exemplified by the in vitro transcription in “nucleus” liposomes and the mimicry of the architecture of eukaryotic cells. Finally, we show the transport of small molecules across the nucleic envelope via insertion of nanopores into the bilayers.
Spatiotemporal organization of distinct biological processes in cytomimetic compartments is a crucial step towards engineering functional artificial cells. Mimicking controlled bi‐directional ...molecular communication inside artificial cells remains a considerable challenge. Here we present photoswitchable molecular transport between programmable membraneless organelle‐like DNA coacervates in a synthetic microcompartment. We use droplet microfluidics to fabricate membraneless non‐fusing DNA coacervates by liquid‐liquid phase separation in a water‐in‐oil droplet, and employ the interior DNA coacervates as artificial organelles to imitate intracellular communication via photo‐regulated uni‐ and bi‐directional transfer of biomolecules. Our results highlight a promising new route to assembly of multicompartment artificial cells with functional networks.
Photoswitchable bi‐directional trafficking of biomolecules between programmable DNA‐based artificial membraneless organelles in a cell‐like microcompartment has been achieved to imitate intracellular communication towards construction of advanced functional artificial cells.
Multicompartment microcapsules, with each compartment protected by a distinct stimuli‐responsive shell for versatile controlled release, are highly desired for developing new‐generation ...microcarriers. Although many multicompartmental microcapsules have been created, most cannot combine different release styles to achieve flexible programmed sequential release. Here, one‐step template synthesis of controllable Trojan‐horse‐like stimuli‐responsive microcapsules is reported with capsule‐in‐capsule structures from microfluidic quadruple emulsions for diverse programmed sequential release. The nested inner and outer capsule compartments can separately encapsulate different contents, while their two stimuli‐responsive hydrogel shells can individually control the content release from each capsule compartment for versatile sequential release. This is demonstrated by using three types of Trojan‐horse‐like stimuli‐responsive microcapsules, with different combinations of release styles for flexible programmed sequential release. The proposed microcapsules provide novel advanced candidates for developing new‐generation microcarriers for diverse, efficient applications.
Trojan‐horse‐like stimuli‐responsive microcapsules with controllable capsule‐in‐capsule structures are created for diverse programmed sequential release. Different combinations of stimuli‐responsive hydrogel shells allow for flexible control of the content release from each capsule compartment for versatile sequential release. The proposed Trojan‐horse‐like stimuli‐responsive microcapsules provide novel advanced candidates for developing new‐generation microcarriers for diverse, efficient applications.
Measuring the novelty of scientific papers is an important research topic. If most subsequent researches of a focal paper only cite itself instead of citing its references as well, this paper could ...be highly disruptive as it may start a new stream of research. However, due to preferential attachment, even if a focal paper is very disruptive, the subsequent works may still cite both the focal paper and some of its highly cited references. To eliminate the noise caused by these highly cited references, we modify the disruption metric and analyze its performance and robustness. The results show that the improved method could better distinguish Nobel prize winning papers from the others. In addition, the resultant ranking is more stable against the highly cited references and random link removal on citation network.
We propose the first least-order Galerkin model of an incompressible flow undergoing two successive supercritical bifurcations of Hopf and pitchfork type. A key enabler is a mean-field consideration ...exploiting the symmetry of the mean flow and the asymmetry of the fluctuation. These symmetries generalize mean-field theory, e.g. no assumption of slow growth rate is needed. The resulting five-dimensional Galerkin model successfully describes the phenomenogram of the fluidic pinball, a two-dimensional wake flow around a cluster of three equidistantly spaced cylinders. The corresponding transition scenario is shown to undergo two successive supercritical bifurcations, namely a Hopf and a pitchfork bifurcation on the way to chaos. The generalized mean-field Galerkin methodology may be employed to describe other transition scenarios.
The compartmentalization of cell-free gene expression systems in liposomes provides an attractive route to the formation of protocells, but these models do not capture the physical (crowded) ...environment found in living systems. Here, we present a microfluidics-based route to produce monodisperse liposomes that can shrink almost 3 orders of magnitude without compromising their stability. We demonstrate that our strategy is compatible with cell-free gene expression and show increased protein production rates in crowded liposome protocells.
Monodisperse Uni- and Multicompartment Liposomes Deng, Nan-Nan; Yelleswarapu, Maaruthy; Huck, Wilhelm T. S
Journal of the American Chemical Society,
06/2016, Volume:
138, Issue:
24
Journal Article
Peer reviewed
Open access
Liposomes are self-assembled phospholipid vesicles with great potential in fields ranging from targeted drug delivery to artificial cells. The formation of liposomes using microfluidic techniques has ...seen considerable progress, but the liposomes formation process itself has not been studied in great detail. As a result, high throughput, high-yielding routes to monodisperse liposomes with multiple compartments have not been demonstrated. Here, we report on a surfactant-assisted microfluidic route to uniform, single bilayer liposomes, ranging from 25 to 190 μm, and with or without multiple inner compartments. The key of our method is the precise control over the developing interfacial energies of complex W/O/W emulsion systems during liposome formation, which is achieved via an additional surfactant in the outer water phase. The liposomes consist of single bilayers, as demonstrated by nanopore formation experiments and confocal fluorescence microscopy, and they can act as compartments for cell-free gene expression. The microfluidic technique can be expanded to create liposomes with a multitude of coupled compartments, opening routes to networks of multistep microreactors.
Division in synthetic cells Chang, Mu-Yueh; Ariyama, Hirotaka; Huck, Wilhelm T. S ...
Chemical Society reviews,
05/2023, Volume:
52, Issue:
1
Journal Article
Peer reviewed
Open access
The bottom-up construction of a living cell using non-living materials represents a grand challenge in science and technology. Reproduction of cells into similar offspring is key to life, and ...therefore, building a synthetic cell that can autonomously divide is one of the most fundamental tasks that need to be achieved in bottom-up synthetic biology. In this review, we summarize the strategies of inducing synthetic division by using physical, chemical, and biological stimuli, and highlight the future challenges to the construction of autonomous synthetic cell division.
In this review, we summarize the strategies of inducing division in synthetic cells by using physical, chemical, and biological stimuli, and highlight the future challenges to the construction of autonomous synthetic cell division.
PDF
