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Photo-Responsive Phase-Separating Fluorescent Molecules for Intracellular Protein Delivery
Yishu Bao1, Hongfei Chen1, Zhiyi Xu1
1Department of Chemistry, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, China.
Angewandte Chemie (International Ed. in English)
|August 30, 2023
Summary
Researchers developed a photo-responsive phase-separating fluorescent molecule (PPFM) for protein delivery. This molecule forms coacervates that transport proteins into cells and disintegrates with light, releasing the payload into the cytoplasm.
Area of Science:
- Biochemistry
- Cell Biology
- Materials Science
Background:
- Cellular membranes pose a significant barrier to exogenous protein delivery.
- Existing protein delivery vehicles often result in endosomal entrapment, limiting cytosolic access.
- Efficient intracellular protein delivery is crucial for various biological and therapeutic applications.
Purpose of the Study:
- To design and characterize a novel photo-responsive phase-separating molecule for efficient protein delivery across cellular membranes.
- To investigate the mechanism of light-triggered payload release from the designed molecular vehicle.
- To demonstrate the utility of this system for delivering peptides and proteins into mammalian cells.
Main Methods:
- Synthesis and characterization of a photo-responsive phase-separating fluorescent molecule (PPFM).
- Investigation of liquid-liquid phase separation (LLPS) behavior of PPFM in aqueous solutions.
- Assessment of PPFM coacervate disintegration kinetics upon photoirradiation (405 nm LED and laser).
- Evaluation of PPFM's capacity to recruit and deliver proteins and peptides into mammalian cells.
- Analysis of payload release into the cytosolic space post-photolysis.
Main Results:
- A novel photo-responsive phase-separating fluorescent molecule (PPFM) was successfully designed (MW 666.8 Da).
- PPFM forms stable fluorescent coacervates via LLPS in aqueous solution.
- Coacervates efficiently recruit diverse peptides and proteins.
- Photoirradiation (405 nm) triggers rapid disintegration of PPFM coacervates within minutes.
- PPFM facilitates the transmembrane delivery of recruited payloads into mammalian cells, with light-induced release into the cytosol.
Conclusions:
- Photo-responsive phase-separating molecules represent a new class of smart materials for biological applications.
- PPFM coacervates serve as effective transmembrane vehicles for intracellular protein delivery.
- Light-triggered photolysis provides precise control over payload release into the cellular cytoplasm.
- This technology offers a promising platform for targeted intracellular delivery of biomolecules.
