A DNA‐Programmed Liposome Fusion Cascade

Löffler, Philipp M. G.; Ries, Oliver; Rabe, Alexander; Okholm, Anders Hauge; Thomsen, Rasmus P.; Kjems, Jørgen; Vogel, Stefan

doi:10.1002/anie.201703243

Cited by 92 publications

(132 citation statements)

References 33 publications

(56 reference statements)

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“…Even geometrically challenging amphiphilic assemblies like cuboid and dumbbell‐shaped hetero‐vesicles can be obtained by aid of DNA origami scaffolds . Likewise, liposomes can be encapsulated inside rigid DNA nanotemplates or rendered to functionally programmed fusion cascades . DNA‐based approaches were also used to prepare sequence‐defined polymers, nanopatterned polymer sheets of various heights or amphiphilic assemblies from semiconducting polymers as a means for creating soft optoelectronic nanostructures .…”

Section: Dna‐based Polymer Materialsmentioning

confidence: 99%

From DNA Nanotechnology to Material Systems Engineering

2019

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Section: Dna‐based Polymer Materialsmentioning

confidence: 99%

From DNA Nanotechnology to Material Systems Engineering

2019

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“…Numerous model systems for membrane fusion have been developed in recent years, all of which have employed a diverse range of molecules as fusogens. Examples of such fusogens include: hydrogen bonding motifs 1 , DNA [2][3][4][5][6][7][8][9][10][11] , PNA [12][13][14] , coiled-coil peptides [15][16][17][18] , and small molecule recognition motifs [19][20][21][22][23] . These systems exhibit varying efficiencies of fusion: some only facilitate hemifusion, or lipid-mixing; whereas others promote full fusion, resulting in content-mixing.…”

mentioning

confidence: 99%

Controlled Peptide-Mediated Vesicle Fusion Assessed by Simultaneous Dual-Colour Time-Lapsed Fluorescence Microscopy

Mora

Boyle

Kolck

et al. 2020

Sci Rep

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We have employed a model system, inspired by SnARe proteins, to facilitate membrane fusion between Giant Unilamellar Vesicles (GUVs) and Large Unilamellar Vesicles (LUVs) under physiological conditions. in this system, two synthetic lipopeptide constructs comprising the coiled-coil heterodimerforming peptides K 4 , (KiAALKe) 4 , or e 4 , (eiAALeK) 4 , a peG spacer of variable length, and a cholesterol moiety to anchor the peptides into the liposome membrane replace the natural SnARe proteins. GUVs are functionalized with one of the lipopeptide constructs and the fusion process is triggered by adding LUVs bearing the complementary lipopeptide. Dual-colour time lapse fluorescence microscopy was used to visualize lipid-and content-mixing. Using conventional confocal microscopy, lipid mixing was observed on the lipid bilayer of individual GUVs. in addition to lipid-mixing, content-mixing assays showed a low efficiency due to clustering of K 4-functionalized LUVs on the GUVs target membranes. We showed that, through the use of the non-ionic surfactant Tween 20, content-mixing between GUVs and LUVs could be improved, meaning this system has the potential to be employed for drug delivery in biological systems.

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“…Growth of GUVs, on the other hand, has already been demonstrated previously [39,40,41] -making the implementation of multiple growth and division cycles an exciting next goal. This could, for instance, be realized by targeted fusion [42] of SUVs composed of the opposite lipid type. The integration of information storage and replication will be yet another important milestone towards the transition from matter to life, or, in other words, towards a synthetic cell which truly deserves its name.…”

Section: Discussionmentioning

confidence: 99%

Protein-free division of giant unilamellar vesicles controlled by enzymatic activity

Dreher

Spatz

Göpfrich

2019

Preprint

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Cell division is one of the hallmarks of life. Success in the bottom-up assembly of synthetic cells will, no doubt, depend on strategies for the controlled autonomous division of protocellular compartments. Here, we describe the protein-free division of giant unilamellar lipid vesicles (GUVs) based on the combination of two physical principles -phase separation and osmosis. We visualize the division process with confocal fluorescence microscopy and derive a conceptual model based on the vesicle geometry. The model successfully predicts the shape transformations over time as well as the time point of the final pinching of the daughter vesicles. Remarkably, we show that two fundamentally distinct yet highly abundant processes -water evaporation and metabolic activity -can both regulate the autonomous division of GUVs. Our work may hint towards mechanisms that governed the division of protocells and adds to the strategic toolbox of bottom-up synthetic biology with its vision of bringing matter to life."Omni cellulae es cellulae." From the point of view of modern science, Raspail's realization from 1825 [1], popularized by Virchow [2], may state the obvious: Every living cell found on Earth today originates from a preexisting living cell. Bottom-up synthetic biology, however, is challenging this paradigm with the vision to create a synthetic cell from scratch [3,4]. Success unquestionably entails that the synthetic cells must have the capacity to produce offspring, making the implementation of synthetic cell division an exciting goal [5,6,7,8]. Over the course of evolution, living cells have developed a sophisticated machinery to divide their compartments in a highly regulated manner. The reconstitution of a minimal set of components from the procaryotic divisome seems to be a promising route towards synthetic cell division. FtsZ [9,6], ESCRT [10,11] and Min proteins [12] led to shape transformations in lipid vesicles, including budding [11] and the formation of tight constrictions [6]. However, reproducible minimal cell division based on common biological mechanisms has not yet been achieved. These challenges leave room for creative approaches, seeking solutions beyond the mimicry of today's biological cells. One exciting strategy is to assemble a division machinery de novo, by designing active, not necessarily protein-based nanomachines. DNA origami structures have been used to shape and remodel lipid vesicles [13,14,15], although active force-generating contractile motors remain a distant goal. A shortcut towards synthetic cell division is the mechanical division of liposomes, which was achieved with microfluidic splitters [16]. While this is not autonomous, it may jump start exciting new directions by circumventing a longstanding challenge. The exploitation of physico-chemical mechanisms, on the other hand, could lead to autonomous division. Noteworthy theoretical and experimental work describes the shape transformations of single-component [17,18,19] as well as phase-separated liposomes [20,21], determined by th...

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A DNA‐Programmed Liposome Fusion Cascade

Cited by 92 publications

References 33 publications

From DNA Nanotechnology to Material Systems Engineering

From DNA Nanotechnology to Material Systems Engineering

Controlled Peptide-Mediated Vesicle Fusion Assessed by Simultaneous Dual-Colour Time-Lapsed Fluorescence Microscopy

Protein-free division of giant unilamellar vesicles controlled by enzymatic activity

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