Multifaceted Cargo Recruitment and Release from Artificial Membraneless Organelles

Abstract

Liquid‐liquid phase separation (LLPS) drives membraneless organelles (MLOs) formation for organizing biomolecules. Artificial MLOs (AMLOs) have been constructed mostly via the LLPS of engineered proteins capable of regulating limited types of biomolecules. Here, leveraging a minimalist AMLO, driven by LLPS of polymer‐oligopeptide hybrids, enrichment, recruitment, and release of multifaceted cargoes are quantitatively shown, including small fluorescent molecules, fluorophore‐containing macromolecules, proteins, DNAs, and RNAs. Cargoes show up to 105‐fold enrichment, whilst recruitment and release are triggered by variations of temperature, pH, and/or ionic strength. Also, the first efficacious, rapid, and reversible control of aggregation‐induced emission with over 30 folds of modulation of overall fluorescence intensity is achieved, by intensifying the aggregation of luminogens in AMLO. The AMLO is a simple yet versatile platform for potential drug delivery and biosensor applications.

Metastable biological matter in liquid phase separation

“…Unique 'stickers-and-spacers' structural feature reported in T-cell receptor signaling pathway [102], nucleophosmin (NPM1) interacting with proteins comprising arginine-rich linear motifs and ribosomal RNA [103] and pair of polySUMO-polySIM interacting multivalent scaffold proteins [104]. Moreover, learning from nature, the unique 'stickers-and-spacers' structural feature of IDPs have also been exploited as building blocks to design bioinspired materials [105][106][107][108][109], which have been reviewed elsewhere [110].…”

Construction of multiphasic membraneless organelles towards spontaneous spatial segregation and directional flow of biochemical reactions

“…We use IDP-mimicking polymeroligopeptide hybrid (IPH) constructs, as scaffold mimetics to form complex membraneless condensates in vitro. An IPH, recently reported by our group, 36 is an engineered multivalent self-interacting hybrid scaffold with well-dened IDP-inspired interaction modules, which demonstrates capability to phase separate and form micro-sized liquid droplets resembling natural MLOs, with the ability to selectively enrich multiple biomolecules 37 and host biochemical reactions. 36 Mimicking the structural features of fused in sarcoma (FUS) protein, a prototypical IDP and a major scaffolding molecule of intracellular MLOs, [38][39][40] the IPH consists of a exible chargeneutral polymer chain, recapitulating the disordered and exible random-coil like nature of IDPs, graed with FUSderived short oligopeptide motifs responsible for weak and reversible interactions, essential for LLPS (Fig.…”

Interaction-Limited Aggregation: Fine-Tuning the Size of pNIPAM Particles by Association with Hydrophobic Ions

“…It can be used to enhance reaction rates by increasing the concentration of reactants (microreactors) and sequestration/isolation of harmful species. Similar schemes can be used for controlled release or water remediation . This process has also been associated with the origin of life .…”