# Exosome-like nanovesicles from acerola for CRISPR-Cas9 ribonucleoprotein delivery to the central nervous system

## TL;DR

Researchers developed a non-invasive method using plant-derived nanovesicles to deliver CRISPR-Cas9 into the brain, enabling targeted gene editing for neurodegenerative diseases.

## Contribution

Acerola-derived exosome-like nanovesicles with GLP2 peptides enable efficient and targeted CRISPR-Cas9 delivery to the central nervous system.

## Key findings

- AELNs form stable complexes with CRISPR-Cas9 ribonucleoproteins and cross the blood-brain barrier.
- Peptide-tagged AELNs enable selective delivery to GLP2-receptor-expressing neurons in the brain.
- Intranasal administration of AELN/RNP complexes successfully edited the C9orf72 gene in vivo.

## Abstract

An aberrant six-base repeat in intron 1 of C9orf72 is the most frequent cause of solitary and familial amyotrophic lateral sclerosis and frontotemporal dementia. This mutation is a potential target for CRISPR/Cas9-based genome editing. However, the blood-brain barrier and limitations of current viral or nanoparticle-based delivery systems to neurons significantly restrict the clinical application of CRISPR-Cas9 in the brain. To address these challenges, we developed a drug delivery system using acerola-derived exosome-like nanoparticles (AELNs), which may overcome several limitations associated with human exosomes. AELNs stably form complexes with ribonucleoproteins (RNPs) comprised of Cas9 proteins and guide RNAs (gRNAs). We improved the delivery efficiency and selectivity of AELN/RNP complexes in GLP2-receptor-expressing neurons by incorporating GLP2 peptides into the AELN/RNP complexes. Intranasal administration of peptide-tagged AELN/RNP complexes in vivo confirmed the successful genome editing of C9orf72, demonstrating the potential of this system for treating neurodegenerative diseases. This study presents a potentially innovative approach for in vivo genome editing using a noninvasive delivery system.

Kuroda and colleagues report that acerola-derived exosome-like nanovesicles efficiently deliver CRISPR-Cas9 into the brain via intranasal administration. This non-invasive strategy enables targeted editing of the ALS-associated C9orf72 gene, highlighting plant-derived vesicles as scalable and safe carriers for future gene therapies in neurodegenerative diseases.