Nature's lessons in design: nanomachines to scaffold, remodel and shape membrane compartments

Beales, Paul A.; Ciani, Barbara; Cleasby, Alexa J.

doi:10.1039/c5cp00480b

Cited by 29 publications

(44 citation statements)

References 191 publications

(288 reference statements)

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“…In the past few decades, scientists and engineers developed methods to generate artificial vesicles, or liposomes, as both model systems to study cell biology and drug carriers to interfere with cell behaviour 2, 3 . The geometry of a liposome defines its physical and chemical properties (fusogenicity, binding affinity to proteins, susceptibility to enzymatic modifications, etc.…”

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confidence: 99%

Placing and shaping liposomes with reconfigurable DNA nanocages

et al. 2017

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The diverse structure and regulated deformation of lipid bilayer membranes are among a cell's most fascinating features. Artificial membrane-bound vesicles, known as liposomes, are versatile tools for modeling biological membranes and delivering foreign objects to cells. To fully mimic the complexity of cell membrane and optimize the efficiency of delivery vesicles, controlling liposome shape (both statically and dynamically) is of utmost importance. Here we report the assembly, arrangement, and remodeling of liposomes with designer geometry: all of which are exquisitely controlled by a set of modular, reconfigurable DNA nanocages. Tubular and toroidal shapes, among others, are transcribed from DNA cages to liposomes with high fidelity, giving rise to membrane curvatures present in cells yet previously difficult to construct in vitro. Moreover, the conformational changes of DNA cages drive membrane fusion and bending with predictable outcomes, opening up opportunities for the systematic study of membrane mechanics.Cells have evolved sophisticated mechanisms to regulate membrane shape and dynamics 1 . In the past few decades, scientists and engineers developed methods to generate artificial vesicles, or liposomes, as both model systems to study cell biology and drug carriers to interfere with cell behaviour 2,3 . The geometry of a liposome defines its physical and chemical properties (fusogenicity, binding affinity to proteins, susceptibility to enzymatic modifications, etc.), and therefore needs to be carefully controlled for each specific study and application. While existing techniques are capable of generating size-defined spherical liposomes and certain aspherical vesicles by means of bottom-up lipid assembly and mechanically distorting membranes 4-10 , design and experiment restraints (for example, lipid * Correspondence to: chenxiang.lin@yale.edu. Author contributions Z.Z. initiated the project, designed and carried out most of the experiments, analyzed the data, and prepared most of the manuscript. Y.Y. performed cryo-EM study and prepared the manuscript. F.P. modeled energy input for membrane fusion and prepared the manuscript. M.L. performed tomography study, analyzed the data, and prepared the manuscript. C.L. initiated the project, designed and supervised the study, interpreted the data, and prepared the manuscript. All authors reviewed and approved the manuscript. Competing financial interestsAuthors declare no competing financial interests. HHS Public Access Author Manuscript Author ManuscriptAuthor ManuscriptAuthor Manuscript composition 11-15 ) often limit a method's adaptability, precision, and programmability. In this work, we take a bioinspired approach (namely DNA nanotemplating) to design, build, and change liposome shapes in a programmable, deterministic manner. Unlike previous methods that rely on trial and error to tune vesicle shape, here we directly programed the geometry of a liposome into its DNA template.Rapid development of DNA-origami technique has led to the construction o...

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Placing and shaping liposomes with reconfigurable DNA nanocages

et al. 2017

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“…Since the introduction of electroformation by Angelova and Dimitrov in 1986 (12), this technique has been widely adopted in the biophysics community for the creation and study of model membranes (35). For instance, the method has been used to investigate the membrane phase behavior (36) and mechanical properties (37) of lipid bilayers.…”

Section: Lipid Mixture Experimentsmentioning

confidence: 99%

Biophysical characterization reveals the similarities of liposomes produced using microfluidics and electroformation

Schaich

Morzy

Sleath

et al. 2019

Preprint

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Giant Unilamellar Vesicles (GUVs) are a versatile tool in many branches of science, including biophysics and synthetic biology. Octanol-Assisted Liposome Assembly (OLA), a recently developed microfluidic technique enables the production and testing of GUVs within a single device under highly controlled experimental conditions. It is therefore gaining significant interest as a platform for use in drug discovery, the production of artificial cells and more generally for controlled studies of the properties of lipid membranes. In this work, we expand the capabilities of the OLA technique by forming GUVs of tunable binary lipid mixtures of DOPC, DOPG and DOPE. Using fluorescence recovery after photobleaching we investigated the lateral diffusion coefficients of lipids in OLA liposomes and found the expected values in the range of 1 μm 2 /s for the lipid systems tested. We studied the OLA derived GUVs under a range of conditions and compared the results with electroformed vesicles. Overall, we found the lateral diffusion coefficients of lipids in vesicles obtained with OLA to be quantitatively similar to those in vesicles obtained via traditional electroformation. Our results provide a quantitative biophysical validation of the quality of OLA derived GUVs, which will facilitate the wider use of this versatile platform.

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“…To emulate this dynamic nature within an artificial cell system, we take inspiration from biology by repurposing protein complexes in vitro that natively remodel membrane structures. 9 The "vesicles-within-a-vesicle" architecture we aspire to recreate in an artificial cell bears resemblance to a cellular membrane-bound organelle, the multivesicular body (MVB). In eukaryotic cells, MVBs are formed by the encapsulation of biomolecular cargo into intraluminal vesicles (ILVs) within endosomes.…”

Section: Figure 1 Experimental Design Rationale Amentioning

confidence: 99%

In vitro membrane remodelling by ESCRT-II/ESCRT-III is regulated by negative feedback from membrane tension

Booth

Ciani

et al. 2018

Preprint

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Artificial cells can shed new light on the molecular basis for life and hold potential for new chemical technologies. Inspired by how nature dynamically regulates its membrane compartments, we aim to repurpose the endosomal sorting complex required for transport (ESCRT) to generate complex membrane architectures as suitable scaffolds for artificial cells.Purified ESCRT-III components perform topological transformations on giant unilamellar vesicles (GUVs) to create complex "vesicles-within-a-vesicle" architectures resembling the compartmentalisation in eukaryotic cells. Thus far, the proposed mechanisms for this activity are based on how assembly and disassembly of ESCRT-III on the membrane drives deformation. Here we demonstrate the existence of a negative feedback mechanism from membrane mechanics that regulates ESCRT-III activity. ILV formation removes excess membrane area, increasing tension, which in turn suppresses downstream ILV formation.This mechanism for in vitro regulation of ESCRT-III activity may also have important implications for its in vivo functions.

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Nature's lessons in design: nanomachines to scaffold, remodel and shape membrane compartments

Cited by 29 publications

References 191 publications

Placing and shaping liposomes with reconfigurable DNA nanocages

Placing and shaping liposomes with reconfigurable DNA nanocages

Biophysical characterization reveals the similarities of liposomes produced using microfluidics and electroformation

In vitro membrane remodelling by ESCRT-II/ESCRT-III is regulated by negative feedback from membrane tension

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