Calcium Shock Enables Efficient and Programmable Particle Delivery for Genome Editing Applications

Area of Science:

• Biotechnology
• Cell Biology
• Molecular Biology

Background:

• Traditional intracellular delivery methods like transfection and transduction show limitations in efficiency, especially with confluent cells and organoids.
• These methods often lack the cell type-specific programmability required for complex biological systems.

Purpose of the Study:

• To introduce and validate a novel methodology, calcium shock (CaSh), for enhanced intracellular particle delivery.
• To develop a programmable version, CaSh-Pro, for targeted delivery into specific cell types within heterogeneous populations.

Main Methods:

• Calcium shock (CaSh) was employed to increase endocytotic uptake and disrupt cell-cell junctions, facilitating particle entry.
• CaSh-Pro integrated molecular targeting agents and amphiphilic peptides for cell-specific delivery and editing.
• The method was tested for delivery of plasmids, ribonucleoproteins, and adeno-associated viral vectors into diverse organoid models.

Main Results:

• CaSh significantly improved particle delivery into single cells, colonies, and organoids.
• CaSh-Pro demonstrated programmable delivery and preferential editing of specific cell types within heterocellular populations.
• The CaSh method showed minimal toxicity and enhanced expression of various genetic materials in intact organoids.

Conclusions:

• CaSh and CaSh-Pro offer simple, versatile protocols for efficient genome editing in complex cellular systems.
• These novel methods hold significant potential for advancing biological discovery and therapeutic development.
• The enhanced delivery and programmability open new avenues for genetic manipulation in diverse cell lineages.