A new architecture for DNA-templated synthesis in which abasic sites protect reactants from degradation

Angewandte Chemie International Edition Wiley 63:14 (2024) e202317482

Authors:

Jennifer Frommer, Robert Oppenheimer, Benjamin M Allott, Samuel Núñez-Pertíñez, Thomas R Wilks, Liam R Cox, Jonathan Bath, Rachel K O'Reilly, Andrew Turberfield

Abstract:

The synthesis of artificial sequence-defined polymers that match and extend the functionality of proteins is an important goal in materials science. One way of achieving this is to program a sequence of chemical reactions between precursor building blocks by means of attached oligonucleotide adapters. However, hydrolysis of the reactive building blocks has so far limited the length and yield of product that can be obtained using DNA-templated reactions. Here, we report an architecture for DNA-templated synthesis in which reactants are tethered at internal abasic sites on opposite strands of a DNA duplex. We show that an abasic site within a DNA duplex can protect a nearby thioester from degradation, significantly increasing the yield of a DNA-templated reaction. This protective effect has the potential to overcome the challenges associated with programmable sequence-controlled synthesis of long non-natural polymers by extending the lifetime of the reactive building blocks.

Coarse-grained modelling of DNA-RNA hybrids

(2023)

Authors:

Eryk J Ratajczyk, Petr Šulc, Andrew J Turberfield, Jonathan PK Doye, Adriaan A Louis

DNA-based optical sensors for forces in cytoskeletal networks

ACS Applied Nano Materials American Chemical Society 6:17 (2023) 15455-15464

Authors:

Christina Jayachandran, Arindam Ghosh, Meenakshi Prabhune, Jonathan Bath, Andrew JJ Turberfield, Lara Hauke, Jorg Enderlein, Florian Rehfeldt, Christoph FF Schmidt

Abstract:

Mechanical forces are relevant for many biological processes, from wound healing and tumor formation to cell migration and differentiation. Cytoskeletal actin is largely responsible for responding to forces and transmitting them in cells, while also maintaining cell shape and integrity. Here, we describe a FRET-based hybrid DNA-protein tension sensor that is designed to sample transient forces in actin networks by employing two actin-binding motifs with a fast off-rate attached to a central DNA hairpin loop. Such a sensor will be useful to monitor rapidly changing stresses in the cell cytoskeleton. We use fluorescence lifetime imaging to determine the FRET efficiency and thereby the conformational state of the sensor, which makes the measurement robust against intensity variations. We demonstrate the applicability of the sensor by confocal microscopy and by monitoring crosslinking activity in in vitro actin networks by bulk rheology.

Designing the self-assembly of arbitrary shapes using minimal complexity building blocks

ACS Nano American Chemical Society 17:6 (2023) 5387-5398

Authors:

Joakim Bohlin, Andrew J Turberfield, Ard A Louis, Petr Šulc

Abstract:

The design space for self-assembled multicomponent objects ranges from a solution in which every building block is unique to one with the minimum number of distinct building blocks that unambiguously define the target structure. We develop a pipeline to explore the design spaces for a set of structures of various sizes and complexities. To understand the implications of the different solutions, we analyze their assembly dynamics using patchy particle simulations and study the influence of the number of distinct building blocks, and the angular and spatial tolerances on their interactions, on the kinetics and yield of the target assembly. We show that the resource-saving solution with a minimum number of distinct blocks can often assemble just as well (or faster) than designs where each building block is unique. We further use our methods to design multifarious structures, where building blocks are shared between different target structures. Finally, we use coarse-grained DNA simulations to investigate the realization of multicomponent shapes using DNA nanostructures as building blocks.

A modular RNA delivery system comprising spherical nucleic acids built on endosome-escaping polymeric nanoparticles

Nanoscale Advances Royal Society of Chemistry (RSC) (2023)

Authors:

Antonio Garcia-Guerra, Ruth Ellerington, Jens Gaitzsch, Jonathan Bath, Mahnseok Kye, Miguel A Varela, Giuseppe Battaglia, Matthew JA Wood, Raquel Manzano, Carlo Rinaldi, Andrew J Turberfield

Abstract:

<jats:p>Polymeric spherical nucleic acids comprise pH-sensitive, polymer-conjugated oligonucleotides that self-assemble into nanoparticles with the ability to escape endosomes, overcoming a major obstacle in nucleic acid delivery.</jats:p>