Near-Infrared Light Activated Formulation for the Spatially Controlled Release of CRISPR-Cas9 Ribonucleoprotein for...

Area of Science:

• Biotechnology
• Neuroscience
• Gene Editing

Background:

• CRISPR/Cas9 gene editing holds therapeutic potential but faces delivery challenges.
• Efficient and safe delivery systems are crucial for clinical translation of gene editing technologies.
• Targeting the brain with gene editing tools requires specialized delivery strategies.

Purpose of the Study:

• To develop and evaluate a novel nanoparticle (NP) formulation for targeted CRISPR/Cas9 gene editing in the brain.
• To demonstrate the efficiency and safety of the NP-based delivery system in vitro and in vivo.
• To enable spatially controlled delivery of gene editing components to specific brain regions.

Main Methods:

• A gene-editing nanoparticle (NP) formulation was designed, immobilizing sgRNA:Cas9 ribonucleoprotein complexes on NP surfaces.
• Near-infrared (NIR) laser irradiation was used to trigger heat generation and release of the ribonucleoprotein complex.
• In vitro gene editing potential was assessed at the single-cell level.
• In vivo safety and gene editing efficacy were evaluated in reporter mice following intracerebral and intranasal administration.

Main Results:

• The NP formulation demonstrated successful gene editing in vitro at the single-cell level.
• In vivo studies in reporter mice confirmed the safety and gene editing capability of the formulation.
• Spatially controlled delivery was achieved in the subventricular zone (intracerebral) and olfactory bulb (intranasal).

Conclusions:

• A rationally designed nanoparticle formulation enables efficient and spatially controlled delivery of CRISPR/Cas9 to the brain.
• The NIR-triggered release system offers a novel strategy for safe and targeted gene editing applications in neurological contexts.
• This advancement paves the way for improved therapeutic strategies for brain disorders using gene editing.