Programmable solid-state condensates for spatiotemporal control of mammalian gene expression.

Area of Science:

• Molecular Biology
• Synthetic Biology
• Biochemistry

Background:

• Precise control over mammalian gene expression is crucial for research and therapeutics.
• Current methods for inducible gene expression often face limitations in specificity and timing.
• Engineering nuclear condensates offers a promising avenue for advanced gene regulation.

Purpose of the Study:

• To develop a versatile strategy for chemically programmed, ligand-dependent gene expression.
• To demonstrate the ability to regulate gene expression at various cellular stages and loci.
• To establish a novel approach for translational control via mRNA retention in nuclear condensates.

Main Methods:

• Harnessing the phase-separation properties of biomolecular condensates.
• Engineering synthetic anchor proteins to tether condensate structures to cellular compartments or gene products.
• Utilizing synthetic solid-state condensates for inducible gene regulation.
• Applying the strategy to both endogenous and episomal gene loci.

Main Results:

• Achieved inducible regulation of transcriptional and translational activities using engineered anchor proteins and synthetic condensates.
• Demonstrated comparable or superior performance to state-of-the-art strategies for CRISPR-Cas9 activity and transcriptional silencing.
• Showcased chemically inducible mRNA retention within nuclear condensates as an efficient translational regulation method.

Conclusions:

• The developed condensate-based strategy provides a versatile and effective platform for precise, on-demand control of mammalian gene expression.
• This approach offers significant advantages over existing methods, particularly in achieving both transcriptional and translational regulation.
• Chemically inducible nuclear condensates represent a powerful tool for advancing synthetic biology and gene therapy applications.