Phenylboronic Acid-Functionalized Polyplexes Tailored to Oral CRISPR Delivery

Area of Science:

• Biotechnology
• Gene Therapy
• Polymer Science

Background:

• CRISPR-Cas9 gene editing holds therapeutic promise, but effective delivery remains a challenge.
• Oral delivery of CRISPR-Cas9 components is largely unexplored due to gastrointestinal tract degradation.
• Developing safe and efficient oral delivery systems is critical for widespread CRISPR application.

Purpose of the Study:

• To develop and evaluate novel oral delivery vehicles for CRISPR-Cas9 components.
• To overcome the limitations of the gastrointestinal environment for gene editing delivery.
• To establish a patient-compliant method for CRISPR-Cas9 administration.

Main Methods:

• Synthesized and characterized novel phenylboronic acid (PBA)-functionalized chitosan-polyethylenimine (CS-PEI) polymers.
• Screened a library of 12 PBA-functionalized CS-PEI polyplexes for optimal properties.
• Evaluated the delivery efficiency, cellular uptake, and gene editing capability in vitro and in vivo (mice).

Main Results:

• Identified an optimized PBA-functionalized CS-PEI formulation demonstrating enhanced mucus penetration and intestinal mucosa targeting.
• The optimized formulation facilitated efficient endosomal escape and cytosolic delivery of CRISPR-Cas9 components.
• Achieved feasible CRISPR-mediated target protein downregulation and subsequent cholesterol reduction in vivo.
• Demonstrated the potential for both local and systemic gene editing via oral administration.

Conclusions:

• Phenylboronic acid-functionalized CS-PEI polymers represent a novel and effective platform for oral CRISPR-Cas9 delivery.
• This approach overcomes key barriers to gastrointestinal delivery, offering a patient-compliant alternative.
• The study highlights a significant advancement towards realizing the therapeutic potential of oral gene editing.