Selective Binding of DNA Origami on Biomimetic Lipid Patches

Hirtz, Michael; Brglez, Josipa; Fuchs, Harald; Niemeyer, Christof M.

doi:10.1002/smll.201501333

Cited by 15 publications

(13 citation statements)

References 38 publications

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“…7 ) and ensured long-term stability of the nanostructures 27 with an imaging buffer containing 5 mM MgCl 2 and 300 mM NaCl in which the Na + outcompetes membrane-adsorbed divalent cations via a counterion release mechanism to break up Mg 2+ promoted interactions between DNA and the phospholipids 28 , 29 . To achieve side-specific binding of the curved DNA origami structures to lipid membranes, we tested various methods including neutravidin-mediated attachment 21 , 30 of biotinylated origami H to biotinylated lipids (Supplementary Fig. 8a ) and covalent attachment 24 , 31 of thiolated origami H to maleimide-modified lipids (Supplementary Fig.…”

Section: Resultsmentioning

confidence: 99%

Membrane sculpting by curved DNA origami scaffolds

et al. 2018

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Membrane sculpting and transformation is essential for many cellular functions, thus being largely regulated by self-assembling and self-organizing protein coats. Their functionality is often encoded by particular spatial structures. Prominent examples are BAR domain proteins, the ‘banana-like’ shapes of which are thought to aid scaffolding and membrane tubulation. To elucidate whether 3D structure can be uncoupled from other functional features of complex scaffolding proteins, we hereby develop curved DNA origami in various shapes and stacking features, following the presumable design features of BAR proteins, and characterize their ability for membrane binding and transformation. We show that dependent on curvature, membrane affinity and surface density, DNA origami coats can indeed reproduce the activity of membrane-sculpting proteins such as BAR, suggesting exciting perspectives for using them in bottom-up approaches towards minimal biomimetic cellular machineries.

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Section: Resultsmentioning

confidence: 99%

Membrane sculpting by curved DNA origami scaffolds

et al. 2018

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“…Similar structures without cholesteryl-TEG anchors appeared to bind only membranes having a positive charge (i.e., with DOTAP), particularly at low salt concentrations, which is expected for a negatively charged polyelectrolyte like DNA ( 51 ). Most recently, the weak selectivity of cholesteryl-anchored origami DNA structures with respect to membranes containing differently charged phospholipids at physiological ionic strength was shown using arrays of supported lipid bilayer patches ( 52 ). Electrostatic DNA-membrane interactions may also be affected by the phase behavior of the membrane.…”

Section: Main Textmentioning

confidence: 99%

DNA Nanostructures on Membranes as Tools for Synthetic Biology

Czogalla

Franquelim

Schwille

2016

Biophysical Journal

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Over the last decade, functionally designed DNA nanostructures applied to lipid membranes prompted important achievements in the fields of biophysics and synthetic biology. Taking advantage of the universal rules for self-assembly of complementary oligonucleotides, DNA has proven to be an extremely versatile biocompatible building material on the nanoscale. The possibility to chemically integrate functional groups into oligonucleotides, most notably with lipophilic anchors, enabled a widespread usage of DNA as a viable alternative to proteins with respect to functional activity on membranes. As described throughout this review, hybrid DNA-lipid nanostructures can mediate events such as vesicle docking and fusion, or selective partitioning of molecules into phase-separated membranes. Moreover, the major benefit of DNA structural constructs, such as DNA tiles and DNA origami, is the reproducibility and simplicity of their design. DNA nanotechnology can produce functional structures with subnanometer precision and allow for a tight control over their biochemical functionality, e.g., interaction partners. DNA-based membrane nanopores and origami structures able to assemble into two-dimensional networks on top of lipid bilayers are recent examples of the manifold of complex devices that can be achieved. In this review, we will shortly present some of the potentially most relevant avenues and accomplishments of membrane-anchored DNA nanostructures for investigating, engineering, and mimicking lipid membrane-related biophysical processes.

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“…Production of SLBs as a model system comprised of either single or multiple component lipids has been done using vesicle fusion [ 2 , 3 , 4 , 5 , 6 , 7 ], Langmuir–Blodgett/Langmuir–Schaefer deposition [ 8 , 9 ]. Scanning probe lithography methods as Dip-Pen Nanolithography [ 10 , 11 , 12 , 13 ] and Polymer Pen Lithography [ 14 ] could also be utilized for the production of lipid bilayer membrane. Vesicle fusion is the most common method on hydrophilic surfaces [ 1 ].…”

Section: Introductionmentioning

confidence: 99%

Lipid Bilayer Membrane in a Silicon Based Micron Sized Cavity Accessed by Atomic Force Microscopy and Electrochemical Impedance Spectroscopy

et al. 2017

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Supported lipid bilayers (SLBs) are widely used in biophysical research to probe the functionality of biological membranes and to provide diagnoses in high throughput drug screening. Formation of SLBs at below phase transition temperature (Tm) has applications in nano-medicine research where low temperature profiles are required. Herein, we report the successful production of SLBs at above—as well as below—the Tm of the lipids in an anisotropically etched, silicon-based micro-cavity. The Si-based cavity walls exhibit controlled temperature which assist in the quick and stable formation of lipid bilayer membranes. Fusion of large unilamellar vesicles was monitored in real time in an aqueous environment inside the Si cavity using atomic force microscopy (AFM), and the lateral organization of the lipid molecules was characterized until the formation of the SLBs. The stability of SLBs produced was also characterized by recording the electrical resistance and the capacitance using electrochemical impedance spectroscopy (EIS). Analysis was done in the frequency regime of 10−2–105 Hz at a signal voltage of 100 mV and giga-ohm sealed impedance was obtained continuously over four days. Finally, the cantilever tip in AFM was utilized to estimate the bilayer thickness and to calculate the rupture force at the interface of the tip and the SLB. We anticipate that a silicon-based, micron-sized cavity has the potential to produce highly-stable SLBs below their Tm. The membranes inside the Si cavity could last for several days and allow robust characterization using AFM or EIS. This could be an excellent platform for nanomedicine experiments that require low operating temperatures.

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Selective Binding of DNA Origami on Biomimetic Lipid Patches

Cited by 15 publications

References 38 publications

Membrane sculpting by curved DNA origami scaffolds

Membrane sculpting by curved DNA origami scaffolds

DNA Nanostructures on Membranes as Tools for Synthetic Biology

Lipid Bilayer Membrane in a Silicon Based Micron Sized Cavity Accessed by Atomic Force Microscopy and Electrochemical Impedance Spectroscopy

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