CRISPR delivery with extracellular vesicles: Promises and challenges.

Area of Science:

• Biotechnology
• Molecular Biology
• Gene Therapy

Background:

• CRISPR gene editing shows promise for genetic disorders, but safe and efficient delivery remains a hurdle.
• Current delivery methods can cause genotoxicity and immunogenicity.
• Extracellular vesicles (EVs) are natural nanoparticles that can transfer genetic material between cells.

Purpose of the Study:

• To review and analyze current strategies for using EVs to deliver CRISPR/Cas9 components.
• To assess the potential and limitations of EV-mediated CRISPR/Cas9 delivery for therapeutic applications.

Main Methods:

• Analysis of various EV-mediated CRISPR/Cas9 delivery strategies.
• Exploration of cell-based, passive loading, active loading, and purified EV loading methods.
• Review of in vitro and in vivo gene editing efficiencies.

Main Results:

• EV-based delivery of CRISPR/Cas9 is effective for both in vitro and in vivo gene editing.
• Multiple strategies exist for loading CRISPR/Cas9 into EVs, including cell-based and direct loading methods.
• Significant variability in cellular uptake and editing efficiency highlights the need for optimization.

Conclusions:

• EVs represent a promising endogenous delivery system for CRISPR/Cas9 gene editing.
• Further standardization and improvement are necessary for therapeutic applications, focusing on purification, loading efficiency, and targeted delivery.