Optogenetic control of RNA function and metabolism using engineered light-switchable RNA-binding proteins.

Area of Science:

• Molecular Biology
• Optogenetics
• RNA Biology

Background:

• RNA-binding proteins (RBPs) are crucial for cellular RNA regulation.
• Current methods for controlling RBP activity in space and time are limited.

Purpose of the Study:

• To engineer a photoswitchable RNA-binding protein (RBP) for optogenetic control.
• To develop a tool for spatiotemporal regulation of RNA functions and gene expression.

Main Methods:

• Engineering of LicV, a blue-light-inducible RBP.
• Fusion of LicV to RNA effectors for controlling RNA localization, splicing, translation, and stability.
• Application of LicV in CRISPR-Cas systems for transcriptional regulation and genomic labeling.

Main Results:

• LicV enables optogenetic control over RNA localization, splicing, translation, and stability in cell culture.
• LicV-assisted CRISPR-Cas systems provide tunable, photoswitchable transcriptional regulation.
• Efficient genomic locus labeling using LicV-based systems was demonstrated.

Conclusions:

• The photoswitchable RBP LicV serves as a programmable scaffold for spatiotemporal control of synthetic RNA effectors.
• LicV offers a novel approach for precise, light-inducible manipulation of RNA biology and gene expression.