Growth and site-specific organization of micron-scale biomolecular devices on living mammalian cells

Abstract

Mesoscale molecular assemblies on the cell surface, such as cilia and filopodia, integrate information, control transport and amplify signals. Designer cell-surface assemblies could control these cellular functions. Such assemblies could be constructed from synthetic components ex vivo, making it possible to form such structures using modern nanoscale self-assembly and fabrication techniques, and then oriented on the cell surface. Here we integrate synthetic devices, micron-scale DNA nanotubes, with mammalian cells by anchoring them by their ends to specific cell surface receptors. These filaments can measure shear stresses between 0-2 dyn/cm2, a regime important for cell signaling. Nanotubes can also grow while anchored to cells, thus acting as dynamic cell components. This approach to cell surface engineering, in which synthetic biomolecular assemblies are organized with existing cellular architecture, could make it possible to build new types of sensors, machines and scaffolds that can interface with, control and measure properties of cells.

Design, Mechanical Properties, and Dynamics of Synthetic DNA Filaments

“…Unpaired filaments were selectively melted away by exchanging the buffer with a solution that contained no free tiles, thus effectively reducing the concentration of the monomer in solution below the critical value necessary for polymer growth. In a recent report, the same group organized micrometer-scale DNA nanotubes at specific receptors on living cells, 83 proposing a model system to mimic the structural and functional role of cell membrane protrusions. For this purpose, the authors engineered a quite complex construct to link DNA nanotubes to epidermal growth factor receptors (EGFR) overexpressed on the surface of HeLa cells (Figure 6c, top panel).…”

“…Confocal micrographs of seeded nanotubes anchored on top of HeLa cell membranes in response to fluid shear stresses of 0, 0.05, 0.2, and 1 dyn/cm 2 . Reprinted with permission from ref . Copyright 2021 Springer Nature.…”

Engineering DNA-based cytoskeletons for synthetic cells

“…The individually polymerizing DNA nanotubes connect over time after reaching a certain length and diffusion time. As a first application DNA origami-seeded DNA nanotubes have been attached on the outer membrane of natural cells to measure the shear flow of the surrounding solution (figure 4 c ) [56]. Inserting the DNA origami seed/DNA nanotube constructs into the membrane of lipid vesicles can even lead to influx of ions and small molecules through the DNA nanotube [59,60].…”

“…( b ) Asymmetrically grown DNA nanotubes connect molecular landmarks [55]. ( c ) DNA nanotubes growing on the cell surface acting as shear force sensors [56]. ( d ) DNA nanotubes growing in angled directions determined by the DNA origami seed conformation [57].…”

DNA‐Programmed Stem Cell Niches via Orthogonal Extracellular Vesicle–Cell Communications

“…The Watson-Crick base pairing of DNA molecules has provided a general approach to program interactions between diverse natural/artificial materials, allowing their precise arrangements from nanometer to millimeter scale. [28][29][30] Particularly, it can mimic natural ligand-receptor recognition, enabling highly predictable and orthogonal interactions between living cells and external interfaces such as nanoparticles, [31,32] liposomes, [33,34] and bulk surfaces. [35,36] Recently, several groups including ours have demonstrated the organization of cell clusters and the modulation of contact cell-cell communications by using DNA strands and/or DNA nanostructures.…”