Smart arginine-equipped polycationic nanoparticles for p/CRISPR delivery into cells

Area of Science:

• Biotechnology
• Nanomedicine
• Gene Therapy

Background:

• Efficient delivery of CRISPR plasmids is crucial for gene editing therapies.
• Existing non-viral vectors often face challenges with transfection efficiency and cellular uptake.

Purpose of the Study:

• To design and evaluate a novel nanocarrier for enhanced CRISPR plasmid delivery.
• To assess the transfection efficiency and gene editing capability of the new nanocarrier across various cell types and in vivo models.

Main Methods:

• Synthesized PEI1.8k-Arg nanoparticles by integrating an arginine-disulfide linker with low-molecular-weight PEI.
• Evaluated nanoparticle complexation with CRISPR plasmid (p/CRISPR).
• Assessed transfection efficiency in HEK 293T, HUVECs, HeLa, and PC-12 cells, and gene editing in HEK 293T-GFP reporter cells.

Main Results:

• PEI1.8k-Arg nanoparticles demonstrated significantly higher transfection efficiency compared to native PEI1.8k.
• The nanocarrier successfully transfected diverse cell types, including hard-to-transfect primary and neuronal cells, with 5-10 times greater efficiency than standard agents.
• Achieved GFP gene editing in HEK 293T-GFP cells using various CRISPR/Cas9 formats and demonstrated successful delivery into brain tissue.

Conclusions:

• PEI1.8k-Arg nanoparticles represent a novel and effective non-viral vector for CRISPR plasmid delivery.
• This nanocarrier system shows broad applicability across different cell types and holds promise for in vivo gene editing applications.
• The developed system is expected to advance gene editing therapies, particularly for treating genetic diseases.