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Directed Protein Packaging within Outer Membrane Vesicles from Escherichia coli: Design, Production and Purification

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Published on: November 16, 2016

A single-component, light-assisted uncaging switch for endoproteolytic release.

Mingguang Cui^1,2, Seunghwan Lee^1,2, Sung Hwan Ban^1,2

¹Department of Anatomy, Korea University College of Medicine, Seoul, Republic of Korea.

Nature Chemical Biology

|November 17, 2023

Summary

This summary is machine-generated.

Researchers developed LAUNCHER, a single-component photocleavable switch using potyviral protease. This system offers precise, user-controlled payload release for synthetic biology applications.

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Area of Science:

Synthetic biology
Molecular biology
Biochemistry

Background:

Proteases act as crucial molecular switches in biological processes.
Potyviral proteases are utilized as reliable proteolytic switches in synthetic biology circuits.

Purpose of the Study:

To develop a single-component photocleavable switch for precise payload release.
To enhance synthetic biological circuits using a novel light-activated system.

Main Methods:

Engineered a circularly permutated tobacco etch virus protease.
Designed a blue-light-gated substrate connected by intermodular linkers.
Integrated the system into a single-component construct termed LAUNCHER (Light-Assisted UNcaging switCH for Endoproteolytic Release).

Main Results:

LAUNCHER demonstrated a superior signal-to-noise ratio compared to multi-component systems.
Achieved precise and user-controllable release of payloads.
Successfully applied LAUNCHER in transgene expression, subcellular translocation, and optochemogenetics.

Conclusions:

LAUNCHER provides a highly efficient and controllable method for payload release in cellular applications.
The plug-and-play nature of LAUNCHER facilitates seamless integration and performance enhancement of synthetic circuits.
LAUNCHER holds significant potential for diverse applications in synthetic biology and beyond.