Thermo-responsive aqueous two-phase system for two-level compartmentalization

Huanqing Cui, Yage Zhang, Sihan Liu, Yang Cao, Qingming Ma, Yuan Liu, Haisong Lin, Chang Li, Yang Xiao, Sammer Ul Hassan and Ho Cheung Shum ()
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Huanqing Cui: The University of Hong Kong
Yage Zhang: Advanced Biomedical Instrumentation Centre
Sihan Liu: The University of Hong Kong
Yang Cao: The University of Hong Kong
Qingming Ma: Qingdao University
Yuan Liu: The University of Hong Kong
Haisong Lin: The University of Hong Kong
Chang Li: The University of Hong Kong
Yang Xiao: The University of Hong Kong
Sammer Ul Hassan: The University of Hong Kong
Ho Cheung Shum: The University of Hong Kong

Nature Communications, 2024, vol. 15, issue 1, 1-13

Abstract: Abstract Hierarchical compartmentalization responding to changes in intracellular and extracellular environments is ubiquitous in living eukaryotic cells but remains a formidable task in synthetic systems. Here we report a two-level compartmentalization approach based on a thermo-responsive aqueous two-phase system (TR-ATPS) comprising poly(N-isopropylacrylamide) (PNIPAM) and dextran (DEX). Liquid membraneless compartments enriched in PNIPAM are phase-separated from the continuous DEX solution via liquid-liquid phase separation at 25 °C and shrink dramatically with small second-level compartments generated at the interface, resembling the structure of colloidosome, by increasing the temperature to 35 °C. The TR-ATPS can store biomolecules, program the spatial distribution of enzymes, and accelerate the overall biochemical reaction efficiency by nearly 7-fold. The TR-ATPS inspires on-demand, stimulus-triggered spatiotemporal enrichment of biomolecules via two-level compartmentalization, creating opportunities in synthetic biology and biochemical engineering.

Date: 2024
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DOI: 10.1038/s41467-024-51043-z

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