Viral Vector-Based Gene Therapy for Hearing Restoration in a Mouse Model

Take an anesthetized mouse pup with genetically induced hearing loss.

In this condition, hair cells in the inner ear lack glutamate transporters, preventing glutamate from being packaged into vesicles and released.

This disrupts the transmission of sound signals to the auditory nerve.

Shave the mouse behind its left ear, disinfect, and apply eye ointment.

Extend the neck and make an incision.

Retract the adipose tissue and muscles to expose the temporal bone.

Perforate the tympanic bulla, part of the temporal bone, and widen the opening to expose the round window membrane or RWM, a thin barrier to the inner ear.

Puncture the RWM and inject the adeno-associated viral vector carrying the therapeutic gene.

The viral particles diffuse and enter hair cells.

Inside hair cells, viral particles deliver their genetic material to the nucleus, triggering glutamate transporter production.

This allows glutamate packaging and release, restoring sound signaling to the auditory nerve.

After confirming anesthesia by toe pinch, shave the left postauricular region and disinfect the skin of a postnatal day 10 to 12 mouse with 70% ethanol and povidone iodine. Then cover the eyes with a protective ophthalmic oint