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Related Experiment Video

Updated: May 21, 2025

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Optogenetic tools for inducing organelle membrane rupture.

Yuto Nagashima1, Tomoya Eguchi1, Ikuko Koyama-Honda1

  • 1Department of Biochemistry and Molecular Biology, Graduate School and Faculty of Medicine, The University of Tokyo, Tokyo, Japan.

The Journal of Biological Chemistry
|March 20, 2025
PubMed
Summary

Researchers developed optogenetic tools to control organelle membrane rupture. This new method, LOV2-BAX, allows for precise, light-activated disruption of mitochondria, ER, and lysosomes, aiding disease research.

Keywords:
Bcl-2-associated X protein (BAX)light-oxygen-voltage-sensing 2 (LOV2) domainlysosomal membrane permeabilization (LMP)membrane rupturemitochondrial outer membrane permeabilization (MOMP)optogenetics

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biotechnology

Background:

  • Organelle membrane disintegration triggers cellular responses and is implicated in diseases like neurodegeneration.
  • Precisely controlling organelle membrane rupture is crucial for studying its effects but remains a significant challenge.
  • Existing methods lack the spatiotemporal control needed to investigate the consequences of specific organelle membrane damage.

Purpose of the Study:

  • To develop novel optogenetic tools for spatiotemporal induction of organelle membrane rupture.
  • To engineer a system that allows researchers to precisely trigger the disintegration of specific organelles using light.
  • To create a versatile platform for studying the cellular and pathological outcomes of organelle membrane damage.

Main Methods:

  • Engineered the Bcl-2-associated X protein (BAX) by replacing its transmembrane domain with organelle-targeting sequences for mitochondria, lysosomes, and endoplasmic reticulum (ER).
  • Fused the photosensitive light-oxygen-voltage-sensing 2 (LOV2) domain to the N-terminus of BAX to create a light-regulatable protein.
  • Utilized blue light to activate the LOV2-BAX fusion protein, inducing membrane rupture in targeted organelles.

Main Results:

  • The engineered BAX mutants successfully targeted and ruptured membranes of mitochondria, lysosomes, and the ER.
  • The LOV2-BAX fusion protein demonstrated blue light-dependent control over organelle membrane rupture.
  • This optogenetic system provides spatiotemporal control over membrane disruption in multiple organelles.

Conclusions:

  • LOV2-BAX is an effective optogenetic tool for inducing organelle membrane rupture with spatiotemporal precision.
  • This technology expands research capabilities for investigating the consequences of organelle membrane damage in various cellular contexts.
  • The developed system offers a novel approach to study autoinflammatory diseases and neurodegeneration linked to organelle dysfunction.