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Instruments / Technical Area	Publication Details	Published
I22-Small angle scattering & Diffraction	Reaction-diffusion patterning of DNA-based artificial cells Adrian Leathers , Michal Walczak , Ryan A. Brady , Assala Al Samad , Jurij Kotar , Michael J. Booth , Pietro Cicuta , Lorenzo Di Michele Journal Of The American Chemical Society 13 DOI: 10.1021/jacs.2c06140 Diamond Proposal Number(s): [28071] Open Access Show Abstract ▼ Abstract: Biological cells display complex internal architectures with distinct micro environments that establish the chemical heterogeneity needed to sustain cellular functions. The continued efforts to create advanced cell mimics, namely, artificial cells, demands strategies for constructing similarly heterogeneous structures with localized functionalities. Here, we introduce a platform for constructing membraneless artificial cells from the self-assembly of synthetic DNA nanostructures in which internal domains can be established thanks to prescribed reaction–diffusion waves. The method, rationalized through numerical modeling, enables the formation of up to five distinct concentric environments in which functional moieties can be localized. As a proof-of-concept, we apply this platform to build DNA-based artificial cells in which a prototypical nucleus synthesizes fluorescent RNA aptamers that then accumulate in a surrounding storage shell, thus demonstrating the spatial segregation of functionalities reminiscent of that observed in biological cells.	Sep 2022
I22-Small angle scattering & Diffraction	Modulating membrane fusion through the design of fusogenic DNA circuits and bilayer composition Miguel Paez-Perez , I. Alasdair Russell , Pietro Cicuta , Lorenzo Di Michele Soft Matter 112 DOI: 10.1039/D2SM00863G Diamond Proposal Number(s): [28071] Open Access Show Abstract ▼ Abstract: Membrane fusion is a ubiquitous phenomenon linked to many biological processes, and represents a crucial step in liposome-based drug delivery strategies. The ability to control, ever more precisely, membrane fusion pathways would thus be highly valuable for next generation nano-medical solutions and, more generally, the design of advanced biomimetic systems such as synthetic cells. In this article, we present fusogenic nanostructures constructed from synthetic DNA which, different from previous solutions, unlock routes for modulating the rate of fusion and making it conditional to the presence of soluble DNA molecules, thus demonstrating how membrane fusion can be controlled through simple DNA-based molecular circuits. We then systematically explore the relationship between lipid-membrane composition, its biophysical properties, and measured fusion efficiency, linking our observations to the stability of transition states in the fusion pathway. Finally, we observe that specific lipid compositions lead to the emergence of complex bilayer architectures in the fusion products, such as nested morphologies, which are accompanied by alterations in biophysical behaviour. Our findings provide multiple, orthogonal strategies to program lipid-membrane fusion, which leverage the design of either the fusogenic DNA constructs or the physico/chemical properties of the membranes, and could thus be valuable in applications where some design parameters are constrained by other factors such as material cost and biocompatibility, as it is often the case in biotechnological applications.	Aug 2022
B21-High Throughput SAXS I22-Small angle scattering & Diffraction	Flexibility defines structure in crystals of amphiphilic DNA nanostars Ryan Brady , Will T. Kaufhold , Nicholas J. Brooks , Vito Foderà , Lorenzo Di Michele Journal Of Physics: Condensed Matter 31 DOI: 10.1088/1361-648X/aaf4a1 Diamond Proposal Number(s): [17271, 16970] Show Abstract ▼ Abstract: DNA nanostructures with programmable shape and interactions can be used as building blocks for the self-assembly of crystalline materials with prescribed nanoscale features, holding a vast technological potential. Structural rigidity and bond directionality have been recognised as key design features for DNA motifs to sustain long-range order in 3D, but the practical challenges associated with prescribing building-block geometry with sufficient accuracy have limited the variety of available designs. We have recently introduced a novel platform for the one-pot preparation of crystalline DNA frameworks supported by a combination of Watson–Crick base pairing and hydrophobic forces (Brady et al 2017 Nano Lett. 17 3276–81). Here we use small angle x-ray scattering and coarse-grained molecular simulations to demonstrate that, as opposed to available all-DNA approaches, amphiphilic motifs do not rely on structural rigidity to support long-range order. Instead, the flexibility of amphiphilic DNA building-blocks is a crucial feature for successful crystallisation.	Feb 2019
I22-Small angle scattering & Diffraction	An amphiphilic-DNA platform for the design of crystalline frameworks with programmable structure and functionality Ryan A. Brady , Nicholas J. Brooks , Vito Foderà , Pietro Cicuta , Lorenzo Di Michele Journal Of The American Chemical Society DOI: 10.1021/jacs.8b09143 Diamond Proposal Number(s): [17271, 19371] Show Abstract ▼ Abstract: The reliable preparation of functional, ordered, nanostructured frameworks would be a game changer for many emerging technologies, from energy storage to nanomedicine. Underpinned by the excellent molecular recognition of nucleic acids, along with their facile synthesis and breadth of available functionalizations, DNA Nanotechnology is widely acknowledged as a prime route for the rational design of nanostructured materials. Yet, the preparation of crystalline DNA frameworks with programmable structure and functionality remains a challenge. Here we demonstrate the potential of simple amphiphilic DNA motifs, dubbed C-stars, as a versatile platform for the design of programmable DNA crystals. In contrast to all-DNA materials, in which structure depends on the precise molecular details of individual build-ing blocks, the self-assembly of C-stars is controlled uniquely by their topology and symmetry. Exploiting this robust self-assembly principle we design a range of topologically identical, but structurally and chemically distinct C-stars that following a one-pot reaction self- assemble into highly porous, functional, crystalline frameworks. Simple design variations allow us to fine-tune the lattice parameter and thus control the partitioning of macromolecules within the frameworks, embed responsive motifs that can induce isothermal disassembly, and include chemical moieties to capture target proteins specifically and reversibly.	Oct 2018
I22-Small angle scattering & Diffraction	Crystallization of amphiphilic DNA C-Stars Ryan A. Brady , Nicholas J. Brooks , Pietro Cicuta , Lorenzo Di Michele Nano Letters DOI: 10.1021/acs.nanolett.7b00980 Diamond Proposal Number(s): [14217, 15030, 15530] Open Access Show Abstract ▼ Abstract: Many emerging technologies require materials with well-defined 3D nanoscale architectures. Production of these structures is currently underpinned by self-assembling amphiphilic macromolecules or engineered all-DNA building blocks. Both these approaches produce restricted ranges of crystal geometries due to synthetic amphiphiles’ simple shape and limited specificity, or the technical difficulties in designing space-filling DNA motifs with targeted shapes. We have overcome these limitations with amphiphilic DNA-nanostructures, or “C-Stars”, that combine the design freedom and facile functionalization of DNA-based materials with robust hydrophobic interactions. C-Stars self-assemble into single crystals exceeding 40 μm in size with lattice parameters exceeding 20 nm.	Apr 2017

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