Nuclease-Resistant L-DNA Tension Probes Enable Long-Term Force Mapping of Single Cells and Cell Consortia.

DNA-based tension probes with precisely programmableforce response provide important insights into cellularmechanosensing. However, their degradability in cell culture limitstheir use for long-term imaging, for instance, when cells migrate,divide, and differentiate. This is a critical limitation for providinginsights into mechanobiology for these longer-term processes. Here,we present DNA-based tension probes that are entirely designedbased on the stereoisomer of biological D-DNA, i.e., L-DNA. Wedemonstrate that L-DNA tension probes are essentially indestructibleby nucleases and provide days-long imaging without significant lossin image quality. We also show their superiority already for shortimaging times commonly used for classical D-DNA tension probes.We showcase the potential of these resilient probes to image minutemovements, and for generating long term force maps of single cellsand of collectively migrating cell populations.

A facile DNA coacervate platform for engineering wetting, engulfment, fusion and transient behavior.

Biomolecular coacervates are emerging models to understand biological systems and important building blocks for designer applications. DNA can be used to build up programmable coacervates, but often the processes and building blocks to make those are only available to specialists. Here, we report a simple approach for the formation of dynamic, multivalency-driven coacervates using long single-stranded DNA homopolymer in combination with a series of palindromic binders to serve as a synthetic coacervate droplet. We reveal details on how the length and sequence of the multivalent binders influence coacervate formation, how to introduce switching and autonomous behavior in reaction circuits, as well as how to engineer wetting, engulfment and fusion in multi-coacervate system. Our simple-to-use model DNA coacervates enhance the understanding of coacervate dynamics, fusion, phase transition mechanisms, and wetting behavior between coacervates, forming a solid foundation for the development of innovative synthetic and programmable coacervates for fundamental studies and applications.

Photocontrolled DNA nanotubes as stiffness tunable matrices for controlling cellular behavior.

Cell behavior is determined by a variety of properties of the extracellular environment like ligand spacing, nanotopography, and matrix stiffness. Matrix stiffness changes occur during many biological processes like wound healing, tumorigenesis, and development. These spatio-temporal dynamic changes in stiffness can cause significant changes in cell morphology, cell signaling, migration, cytoskeleton etc. In this paper, we have created photocontrolled stiffness-tunable DNA nanotubes which can undergo reversible changes in their conformation upon UV and VIS irradiation. When used as a substrate for cell culture, the photocontrolled DNA nanotubes can tune the cell morphology of HeLa cells from a long spindle-shaped morphology with long filopodia protrusions to a round morphology with short filopodia-like extrusions. Such a photocontrolled nanosystem can give us deep insights into the cell-matrix interactions in the native extracellular matrix caused by nanoscopic changes in stiffness.

Biomimetic DNA Nanotechnology to Understand and Control Cellular Responses.

At the cellular level, numerous nanocues guide the cells to adhere, interact, proliferate, differentiate, etc. Understanding and manipulating the cellular functions in vitro, necessitates the elucidation of these nanocues provided to the cells by the extracellular matrix (ECM), neighbouring cells or in the form of ligands. DNA nanotechnology is a biocompatible, flexible and a promising molecular level toolkit for mimicking cell-cell and cell-matrix interactions. In this review, we summarize various advances in cell-matrix, cell-cell and cell receptor-ligand interactions using DNA nanotechnology as a tool. We also provide a brief outlook on the current challenges and the future potentials of these DNA-based nanostructures so as to inspire novel innovations in the field.

Non-invasive Regulation of Cellular Morphology Using a Photoswitchable Mechanical DNA Polymer.

The extracellular matrix (ECM) in which the cells reside provides a dynamic and reversible environment. Spatiotemporal cues are essential when cells are undergoing morphogenesis, repair and differentiation. Emulation of such an intricate system with reversible presentation of nanoscale cues can help us better understand cellular processes and can allow the precise manipulation of cell function in vitro. Herein, we formulated a photoswitchable DNA mechanical nanostructure containing azobenzene moieties and dynamically regulated the spatial distance between adhesion peptides using a photoswitchable DNA polymer with photoirradiation. We found that the DNA polymer reversibly forms two different structures, a relaxed linear and shrunken compact form, observed by AFM. Using the mechanical properties of this DNA polymer, UV and visible light irradiation induced a significant morphology change of the cells between a round shape and spindle shape, thus providing a tool to decipher the language of the ECM better.