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Near-Infrared Optogenetic Module for Conditional Protein Splicing.

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This study introduces a novel near-infrared optogenetic tool for controlling protein splicing. This new method enables precise light-triggered protein cleavage, offering advanced control over molecular processes.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Optogenetics

Background:

  • Optogenetics offers precise spatiotemporal control of biological processes.
  • Near-infrared (NIR) light is advantageous due to low phototoxicity and deep tissue penetration.
  • Optogenetic control of polypeptide bond formation remains underdeveloped.

Purpose of the Study:

  • To develop a NIR optogenetic module for conditional protein splicing (CPS).
  • To engineer a NIR CPS gene expression system for light-triggered protein cleavage.
  • To demonstrate the application of NIR CPS in activating pyroptotic cell death.

Main Methods:

  • Developed a NIR optogenetic module based on the gp41-1 intein.
  • Optimized the module for minimal background signal and high light/dark contrast.
  • Engineered a NIR CPS gene expression system utilizing protein ligation of a transcription factor.
  • Applied NIR CPS to trigger protein cleavage and activate gasdermin D.

Main Results:

  • Successfully created and optimized a NIR optogenetic module for CPS.
  • Established a functional NIR CPS gene expression system.
  • Demonstrated light-triggered activation of gasdermin D, inducing pyroptotic cell death.
  • Achieved precise control over molecular processes via covalent protein linkage and cleavage.

Conclusions:

  • The developed NIR CPS optogenetic module provides a powerful new tool.
  • This technology enables spatiotemporal control of molecular processes through protein linkage and cleavage.
  • The system holds promise for applications in biotechnology and molecular biology research.