Electroactive nanoinjection platform for intracellular delivery and gene silencing

Area of Science:

• Biotechnology
• Cell Biology
• Nanotechnology

Background:

• Nanoinjection uses nanostructures for efficient intracellular delivery, overcoming limitations of viral carriers like safety and low cargo capacity.
• Existing nanoinjection methods face challenges in precise cellular access and maintaining cell integrity.
• Conventional electroporation requires high voltages, leading to significant cell damage and mortality.

Purpose of the Study:

• To develop a non-viral, low-voltage electroactive nanoinjection (ENI) platform for precise intracellular delivery.
• To enable efficient delivery of diverse biomolecular cargos into cells with minimal perturbation.
• To achieve effective gene silencing using the ENI platform.

Main Methods:

• Development of a reusable electroactive nanoinjection (ENI) platform using vertically configured conductive nanotubes (NTs).
• Utilizing localized electric fields at the NT-cell membrane interface to reduce required voltage to ≤10 V.
• Employing theoretical simulations and experimental validation for platform assessment.

Main Results:

• The ENI platform successfully perforated GPE-86 mouse fibroblast cells, enabling efficient delivery of antibodies (62.5%), mRNA (55.5%), and plasmid DNA (51.8%).
• Cell viability remained high (>90%) post-nanoscale electroporation (EP).
• Demonstrated gene silencing efficacy of 41.3% through siRNA delivery targeting TRIOBP.

Conclusions:

• The developed non-viral, low-voltage ENI platform provides a safe and versatile new method for intracellular delivery.
• This technology facilitates broader cargo and cell type selection for ex vivo cell engineering.
• ENI opens new avenues for advanced gene silencing applications.