2. Academic Validation
  3. Light-mediated intracellular polymerization

Light-mediated intracellular polymerization

  • Nat Protoc. 2024 Mar 21. doi: 10.1038/s41596-024-00970-8.
Mohamed Abdelrahim 1,  Quan Gao 1,  Yichuan Zhang 1 2,  Weishuo Li 3,  Qi Xing 1,  Mark Bradley 4,  Jin Geng 5
Affiliations

Affiliations

  • 1 Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.
  • 2 School of Pharmacy, Henan University, Kaifeng, China.
  • 3 Center for Molecular Metabolism, School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing, China.
  • 4 Precision Healthcare University Research Institute, Queen Mary University of London, London, UK. m.bradley@qmul.ac.uk.
  • 5 Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China. jin.geng@siat.ac.cn.
PMID: 38514838 DOI: 10.1038/s41596-024-00970-8
Abstract

The synthesis of synthetic intracellular Polymers offers groundbreaking possibilities in cellular biology and medical research, allowing for novel experiments in drug delivery, bioimaging and targeted Cancer therapies. These macromolecules, composed of biocompatible monomers, are pivotal in manipulating cellular functions and pathways due to their bioavailability, cytocompatibility and distinct chemical properties. This protocol details two innovative methods for intracellular polymerization. The first one uses 2-hydroxy-4'-(2-hydroxyethoxy)-2-methylpropiophenone (Irgacure 2959) as a photoinitiator for free radical polymerization under UV light (365 nm, 5 mW/cm2). The second method employs photoinduced electron transfer-reversible addition-fragmentation chain-transfer polymerization with visible light (470 nm, 100 mW/cm2). We further elaborate on isolating these intracellular Polymers by streptavidin/biotin interaction or immobilized metal ion affinity chromatography for Polymers tagged with biotin or histidine. The entire process, from polymerization to isolation, takes ~48 h. Moreover, the intracellular Polymers thus generated demonstrate significant potential in enhancing actin polymerization, in bioimaging applications and as a novel avenue in Cancer treatment strategies. The protocol extends to animal models, providing a comprehensive approach from cellular to systemic applications. Users are advised to have a basic understanding of organic synthesis and Cell Biology techniques.

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