Controlling Circular RNA Encapsulation within Extracellular Vesicles for Gene Editing and Protein Replacement

Area of Science:

• Biotechnology
• Molecular Biology
• Cell Biology

Background:

• Extracellular vesicles (EVs) are natural carriers of biomolecules, including RNAs.
• Efficiently loading long RNAs, such as circular RNAs (circRNAs), into EVs is challenging with conventional methods.
• Existing methods may compromise EV integrity or RNA cargo.

Purpose of the Study:

• To develop a novel strategy for high-efficiency encapsulation of long circRNAs into EVs.
• To leverage cellular sorting mechanisms for improved circRNA loading.
• To demonstrate the therapeutic potential of EV-delivered circRNAs.

Main Methods:

• Engineered cells to increase cytoplasmic concentration of target circRNAs.
• Utilized inherent circRNA enrichment in EVs and RNA back-splicing.
• Harnessed cellular sorting pathways for circRNA encapsulation into EVs.

Main Results:

• Achieved highly efficient loading of long circRNAs (>1000 nt) into EVs.
• Demonstrated successful delivery of EV-encapsulated circRNAs to recipient cells.
• Showcased gene editing applications in mice using these engineered EVs.

Conclusions:

• The developed strategy enables efficient encapsulation and delivery of long circRNAs via EVs.
• This method overcomes limitations of conventional RNA loading techniques.
• EV-mediated delivery of circRNAs holds significant promise for gene therapy and protein expression applications.