A large, square-shaped, DNA origami nanopore with sealing function on a giant vesicle membrane

Iwabuchi, Shoji; Kawamata, Ibuki; Murata, Satoshi; Nomura, Sakashi

doi:10.1039/d0cc07412h

Cited by 25 publications

(33 citation statements)

References 41 publications

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“…Inspired by biological nanopores such as αHL, there has been significant advancement in the field of synthetic nanopores, especially ones designed with DNA [70,71]. Unlike αHL, the size of DNA-based nanopores can be tuned from 4 to 30 nm in diameter [72][73][74], and these nanopores have been applied in numerous sensing applications [75].…”

Section: Alpha Hemolysinmentioning

confidence: 99%

Synthetic Cell as a Platform for Understanding Membrane-Membrane Interactions

et al. 2021

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In the pursuit of understanding life, model membranes made of phospholipids were envisaged decades ago as a platform for the bottom-up study of biological processes. Micron-sized lipid vesicles have gained great acceptance as their bilayer membrane resembles the natural cell membrane. Important biological events involving membranes, such as membrane protein insertion, membrane fusion, and intercellular communication, will be highlighted in this review with recent research updates. We will first review different lipid bilayer platforms used for incorporation of integral membrane proteins and challenges associated with their functional reconstitution. We next discuss different methods for reconstitution of membrane fusion and compare their fusion efficiency. Lastly, we will highlight the importance and challenges of intercellular communication between synthetic cells and synthetic cells-to-natural cells. We will summarize the review by highlighting the challenges and opportunities associated with studying membrane–membrane interactions and possible future research directions.

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Section: Alpha Hemolysinmentioning

confidence: 99%

Synthetic Cell as a Platform for Understanding Membrane-Membrane Interactions

et al. 2021

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“…DNA nanopores with inner diameters over 3 nm have been shown to mediate the passage of large biomolecules, such as double-stranded DNA and proteins, across lipid bilayer membranes. [18][19][20][21][22] Moreover, the highly predictable DNA interactions are the basis for the creation of programmable DNA nanopores that initiate transport only in the presence of specific chemical or spatial cues. 20,23 Advancements in DNA modifications with various chemical groups and aptamers and control over pore geometries have further enabled specificities in the selective transport of solute species across the DNA nanopores.…”

Section: Introductionmentioning

confidence: 99%

“…20,23 Advancements in DNA modifications with various chemical groups and aptamers and control over pore geometries have further enabled specificities in the selective transport of solute species across the DNA nanopores. 21,24 In addition to the potential applications in biosensing and drug delivery, 25 DNA nanopores can serve essential functions as transporters in systems of synthetic cells 26 : The lack of specificity of many types of membrane pores and the lack of reliable transport mechanisms between synthetic cells are central challenges in reconstituting complex signaling systems in synthetic cells. 27 The programmability and specificity of DNA nanopores would allow delivery of target genetic and signaling materials to make efficient communications possible.…”

Section: Introductionmentioning

confidence: 99%

Lossless end-to-end transport of small molecules through micron-length DNA nanochannels

Joshi

Aksimentiev

et al. 2022

Preprint

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Designed and engineered protein and DNA nanopores can sense and characterize single molecules and control transmembrane transport of molecular species. However, designed biomolecular pores are less than 100 nm in length and are used primarily for transport across lipid membranes. Nanochannels that span longer distances could be used as conduits for molecules between non-adjacent compartments or cells. Here, we design microns-long, 7 nm diameter DNA nanochannels that small molecules can traverse according to the laws of continuum diffusion. Binding DNA origami caps to channel ends eliminates transport and demonstrates that molecules diffuse from one channel end to the other rather than permeating through channel walls. These micron-length nanochannels can also grow, form interconnects, and interface with living cells. This work thus shows how to construct multifunctional, dynamic agents that control molecular transport, opening new ways of studying intercellular signaling and modulating molecular transport between synthetic and living cells.

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“…In the last decade, advancements in the field of DNA nanotechnology have enabled the fabrication of a great variety of ‘DNA origami’ nanostructures 1,2 , including transmembrane structures that resemble the biological pores found in cells 3 . Drawing inspiration from protein pores such as alpha-hemolysin 4 , artificial pores have been engineered with DNA origami 1,2 that can insert into lipid bilayers and allow for transmembrane diffusion of ions 5,6 and small molecules such as fluorophores 7,8 , DNA oligomers 5,7 , short PEG 9 , and dextran 8,10 . To favor partitioning into the lipid bilayer, the usual strategy relies on the chemical modification of the outer nanopore surface with hydrophobic groups, e.g.…”

Section: Introductionmentioning

confidence: 99%

“…However, so far only small pores with an inner diameter of a few nm have been realized 8,10 , which is because wider pores are increasingly difficult to insert into a membrane. A commonly used strategy to reconstitute pores into a bilayer relies on spontaneous insertion into a preformed lipid membrane 3 .…”

Section: Introductionmentioning

confidence: 99%

Reconstitution of ultrawide DNA origami pores in liposomes for transmembrane transport of macromolecules

Fragasso

Franceschi

Stoemmer

et al. 2021

Preprint

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Molecular traffic across lipid membranes is a vital process in cell biology that involves specialized biological pores with a great variety of pore diameters, from fractions of a nanometer to >30 nm. Creating artificial membrane pores covering similar size and complexity will aid the understanding of transmembrane molecular transport in cells, while artificial pores are also a necessary ingredient for synthetic cells. Here, we report the construction of DNA origami nanopores that have an inner diameter as large as 30 nm. We developed new methods to successfully insert these ultrawide pores into the lipid membrane of giant unilamellar vesicles (GUVs) by administering the pores concomitantly with vesicle formation in an inverted-emulsion cDICE technique. The reconstituted pores permit the transmembrane diffusion of large macromolecules such as folded proteins, which demonstrates the formation of large membrane-spanning open pores. The pores are size selective as dextran molecules with a diameter up to 22 nm can traverse the pores, whereas larger dextran molecules are blocked. By FRAP measurements and modelling of the GFP influx rate, we find that up to hundreds of pores can be functionally reconstituted into a single GUV. Our technique bears great potential for applications across different fields from biomimetics, synthetic biology, to drug delivery.

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A large, square-shaped, DNA origami nanopore with sealing function on a giant vesicle membrane

Abstract: Intaking molecular information from the external environment is essential for the proper functioning of artificial cells/molecular robots. Herein, we report the design and function of a membrane nanopore using a...

Cited by 25 publications

References 41 publications

Synthetic Cell as a Platform for Understanding Membrane-Membrane Interactions

Synthetic Cell as a Platform for Understanding Membrane-Membrane Interactions

Lossless end-to-end transport of small molecules through micron-length DNA nanochannels

Reconstitution of ultrawide DNA origami pores in liposomes for transmembrane transport of macromolecules

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