Spatially-Resolved Organoid Transfection by Porous Silicon-Mediated Optoporation

Area of Science:

• Biotechnology
• Materials Science
• Cell Biology

Background:

• Organotypic cultures are crucial for studying complex biological processes and developing therapies.
• Spatially-controlled gene delivery is essential for creating advanced tissue models.
• Current methods for localized optoporation in 3D systems face significant challenges.

Purpose of the Study:

• To develop a safe and effective method for spatially-resolved nucleic acid delivery in 3D organoid models.
• To utilize porous silicon nanoparticles for optoporation and mRNA transfection.
• To overcome limitations in localized optoporation within complex 3D cellular environments.

Main Methods:

• Porous silicon nanoparticles functionalized with an azobenzene-lysine photo-switchable moiety were synthesized.
• Near-infrared two-photon optoporation was employed for spatially-resolved transfection.
• Messenger RNA (mRNA) delivery was assessed in MCF-7 organoids and spheroids.

Main Results:

• Porous silicon nanoparticles demonstrated safe and bioresorbable properties.
• Functionalized nanoparticles achieved up to 84% transfection efficiency in 2D cell systems.
• Spatially selective mRNA transfection was successfully achieved in complex 3D MCF-7 spheroids.

Conclusions:

• Porous silicon nanoparticles offer a viable solution for localized 3D optoporation.
• This approach enables targeted gene delivery in complex 3D cellular environments.
• The method provides a pathway for designing tailored spheroids and organoids through precise nucleic acid delivery.