A DNA‐Micropatterned Surface for Propagating Biomolecular Signals by Positional on‐off Assembly of Catalytic Nanocompartments

Abstract

Signal transduction is pivotal for the transfer of information between and within living cells. The composition and spatial organization of specified compartments are key to propagating soluble signals. Here, a high‐throughput platform mimicking multistep signal transduction which is based on a geometrically defined array of immobilized catalytic nanocompartments (CNCs) that consist of distinct polymeric nanoassemblies encapsulating enzymes and DNA or enzymes alone is presented. The dual role of single entities or tandem CNCs in providing confined but communicating spaces for complex metabolic reactions and in protecting encapsulated compounds from denaturation is explored. To support a controlled spatial organization of CNCs, CNCs are patterned by means of DNA hybridization to a microprinted glass surface. Specifically, CNC‐functionalized DNA microarrays are produced where individual reaction compartments are kept in close proximity by a distinct geometrical arrangement to promote effective communication. Besides a remarkable versatility and robustness, the most prominent feature of this platform is the reversibility of DNA‐mediated CNC‐anchoring which renders it reusable. Micropatterns of polymer‐based nanocompartment assemblies offer an ideal scaffold for the development of the next generation responsive and communicative soft‐matter analytical devices for applications in catalysis and medicine.

Polymeric Giant Unilamellar Vesicles with Integrated DNA‐Origami Nanopores: An Efficient Platform for Tuning Bioreaction Dynamics Through Controlled Molecular Diffusion

“…The dispersity (Ð = 1.49) and the number-average molar mass (M n = 1800 g mol −1 ) were determined by gel permeation chromatography, as recently presented elsewhere. [61] DoNs Characterization and Hybridization: The DoNs structure and purity were characterized by the provider (Tilibit Nanosystem) using a combination of transmission electron microscopy (TEM) (Figure S12, Supporting Information) and gel electrophoresis (Figure S13, Supporting Information) experiments. The results indicated the absence of staple strands used for the DoNs synthesis, as well as the absence of DoNs aggregates after filtration and purification.…”

“…The dispersity (Ð = 1.49) and the number‐average molar mass (M n = 1800 g mol −1 ) were determined by gel permeation chromatography, as recently presented elsewhere. [ 61 ]…”

Synthetic Cells Revisited: Artificial Cell Construction Using Polymeric Building Blocks

“…[107] Of particular interest was to use strain-promoted azide-alkyne cycloaddition (SPAAC) click crosslinking to attach linkers, such as ssDNA, on preformed polymersomes which then favor the clustering process, and for fabrication of artificial organelles facilitating spatially controlled enzymatic reactions. [81,108] This approach avoids DNA prefunctionalization of the block copolymers, which would alter the ratio between hydrophilic and hydrophobic blocks and subsequently interfere with the self-assembly process.…”

Advancing the Design of Artificial Nano-organelles for Targeted Cellular Detoxification of Reactive Oxygen Species

“…This can be accomplished by a robust shell that allows enzymes to function in situ while being protected against a harmful environment. , A particularly appealing strategy to enable efficient intracellular ROS detoxification with high spatiotemporal precision involves the development of artificial organelles as counterparts of natural organelles. The advantages of artificial organelles are their tuning to a specific functionality required for the treatment of a pathologic condition and the protection they offer to sensitive molecules inside. − Artificial organelles (AnOs) coencapsulating two detoxifying enzymes inside polymersomes were shown to mimic native peroxisomes in the conversion of ROS, while simpler organelles containing only one type of enzyme were reported to degrade H 2 O 2 in zebra fish embryos . However, the specific cell-targeting by AnOs and their subsequent uptake remain a challenge.…”