ABA-Induced Cargo Proteins Loading in Extracellular Vesicles for Gene Editing

Area of Science:

• Biotechnology
• Molecular Biology
• Drug Delivery Systems

Background:

• Extracellular vesicles (EVs) are promising drug delivery vehicles due to their biocompatibility and targeting abilities.
• Controlled loading of specific proteins into EVs remains a challenge for therapeutic applications.

Purpose of the Study:

• To develop a novel system for controllable protein loading into EVs using an abscisic acid (ABA)-inducible proximity approach.
• To engineer exosomal scaffolds for targeted cargo encapsulation during EV biogenesis.

Main Methods:

• Engineered exosomal scaffolds by fusing the ABA receptor PYL1 to EV-enriched proteins (BASP1, CD9, PTGFRN).
• Tagged target proteins (EGFP, luciferase, Cas9) with the ABI1 phosphatase domain.
• Administered ABA to producer cells to induce PYL1-ABI1 complex formation and cargo recruitment into EVs.

Main Results:

• Demonstrated ABA-inducible and selective encapsulation of cargo proteins into EVs.
• BASP1-PYL1 scaffold showed the most effective ABA-dependent cargo enrichment.
• Purified EVs retained normal morphology, size, and marker expression.
• Delivered functional cargo, including Cas9, to recipient cells for genome editing.

Conclusions:

• Established an ABA-triggered molecular switch for controllable EV protein loading.
• The developed system provides a versatile platform for next-generation therapeutic delivery via engineered EVs.