Expression of nano-engineered RNA organelles in bacteria

Area of Science:

• Synthetic Biology
• Biotechnology
• RNA Nanotechnology

Background:

• Membraneless organelles are crucial cellular components.
• Designing synthetic versions offers insights into natural functions and engineering applications.
• E. coli is a relevant host for biotechnological advancements.

Purpose of the Study:

• To design and express novel synthetic membraneless organelles in vivo using E. coli.
• To achieve precise control over condensate formation, protein recruitment, and dissolution.
• To explore the potential of RNA nanotechnology for cellular and metabolic engineering.

Main Methods:

• Utilized RNA nanotechnology to create designer condensates in E. coli.
• Employed branched RNA motifs for co-transcriptional assembly via base-pairing.
• Incorporated protein-binding aptamers for selective protein recruitment.
• Implemented thermal cycling for reversible condensate dissolution and reassembly.

Main Results:

• Successfully designed and expressed robust, stable synthetic membraneless organelles in E. coli.
• Achieved orthogonal, non-mixing condensates with selective protein recruitment capabilities.
• Demonstrated reversible control over protein client release and recapture.
• Showcased algorithmic control over interactions and condensate microstructure.

Conclusions:

• Nanostructured RNA motifs provide advanced control over synthetic organelles compared to peptide or repetitive RNA systems.
• These designer condensates hold significant potential for synthetic biology and biotechnology.
• The developed system offers a versatile platform for cellular engineering and metabolic control.