Ultrasound-Actuated Gene Editing in Human Kidney Organoids

Area of Science:

• Biotechnology
• Gene Editing
• Nanotechnology

Background:

• Efficient delivery of gene editing ribonucleoproteins (RNPs) into solid tissues is a major challenge for CRISPR-Cas9 clinical applications.
• Non-viral delivery methods often struggle with penetration and distribution within complex 3D cellular structures.

Purpose of the Study:

• To develop and evaluate acoustically-actuated peptide nanoemulsions (NPeps) for enhanced RNP delivery into 3D tissues.
• To assess the efficacy and spatial distribution of NPep-mediated gene editing in human kidney organoids.

Main Methods:

• Development of peptide nanoemulsions (NPeps) capable of being guided and activated by ultrasound.
• Utilizing ultrasound to trigger ballistic delivery of RNPs encapsulated within NPeps into human kidney organoids.
• Comparison of NPep delivery with commercial lipofection reagents regarding editing efficiency and tissue integrity.

Main Results:

• NPeps successfully delivered RNPs into cells within the bulk of dense 3D cellular structures (human kidney organoids).
• NPep vectors demonstrated improved spatial distribution of gene editing compared to lipofection reagents.
• The NPep approach did not disrupt the tissue structure or compromise cell viability.

Conclusions:

• Acoustically-actuated NPeps provide a promising technology for spatiotemporally controlled, deep-tissue RNP delivery.
• This method has the potential to advance imaging-guided CRISPR-Cas9 delivery for diagnostics and therapeutics.
• NPeps represent a significant step towards overcoming barriers in the clinical translation of non-viral gene editing strategies.