Extracellular vesicle–mediated gene editing for the treatment of nonsyndromic progressive hearing loss in adult mice

Nov 12, 2025Science translational medicine

Gene editing delivered by tiny particles to treat progressive hearing loss in adult mice

AI simplified

CRISPR Gene Editing on OpenScience ↗PubMed ↗DOI ↗

Abstract

A 10-fold enhancement in loading efficiency for CRISPR-Cas9 ribonucleoprotein complexes into extracellular vesicles was achieved using a microfluidic droplet-based system.

A novel delivery method utilizing extracellular vesicles (EVs) for in vivo gene therapy was developed.
The microfluidic droplet-based electroporation system (μDES) allowed for more than 1000-fold increase in processing throughput compared to traditional methods.
Ribonucleoprotein complexes were effectively delivered into cochlear hair cells in a mouse model of progressive hearing loss.
Injection of RNP-EVs into the inner ear reduced messenger RNA expression related to hearing loss.
Auditory brainstem responses indicated evidence of hearing preservation in treated mice compared to untreated and EV-only injected controls.

AI simplified

The clinical translation of gene therapy has been challenging in part because of the limitations of current delivery approaches. Herein, we report an efficient nonviral genome editor delivery approach using extracellular vesicles (EVs) carrying single-guide RNA (sgRNA): CRISPR-Cas9 ribonucleoprotein (RNP) complexes for in vivo gene therapy. By leveraging a high-throughput microfluidic droplet-based electroporation system (μDES), we achieved a 10-fold enhancement in loading efficiency and more than 1000-fold increase in processing throughput for loading RNP complexes into EVs compared with conventional high-voltage pulsed electroporation. μDES generated uniform microdroplets containing EVs and RNPs by applying direct current-controlled low voltage (up to 60 V) to transiently permeabilize membranes and enable efficient cargo encapsulation while maintaining EV integrity at both the protein and morphological levels. In themouse model of autosomal dominant progressive hearing loss, which may model MYO7A-associated DFNA11 hearing loss in humans, we demonstrated the effective delivery of RNPs by EVs into cochlear hair cells by cross-sectional and whole-mount confocal imaging. The injection of RNP-EVs via the posterior semicircular canal in 4-week-oldmice resulted in a reduction inmessenger RNA expression and evidence of hearing preservation, as measured by auditory brainstem responses, compared with untreated ears and EV only-injected mice. This study highlights the potential of μDES-produced RNP-EVs for gene editing as a treatment for progressive nonsyndromic hearing loss in patients. Myo7a WT/Sh1Myo7a WT/Sh1Myo7a Sh1

Full Text

Full text is available at the source.

View full text ↗

Extracellular vesicle–mediated gene editing for the treatment of nonsyndromic progressive hearing loss in adult mice

Abstract

Full Text

You found one interesting study. We’ll send the next 7.

what lands in your inbox each week:

Recent issues from the crispr gene editing brief

Abstract

Full Text

Related papers

You found one interesting study. We’ll send the next 7.

what lands in your inbox each week:

Recent issues from the crispr gene editing brief