Intrathecal Delivery of Viruses in a Mouse Model for Treating Central Nervous System Diseases

Fill a syringe with a solution containing adeno-associated viruses, or AAV, and lidocaine, a local anesthetic.

Place an awake mouse in a prone position and cover its upper body.

Hold the mouse's pelvic girdle and shave its lower back to expose the skin. Sterilize it with iodine and ethanol.

Locate the gap in the lower spine and gently press to mark it.

Rotate the tail to align the spine and insert the syringe vertically.

Adjust the angle until a tail flick occurs, confirming needle entry into the intrathecal space where the cerebrospinal fluid or CSF flows.

Slowly inject the solution for even distribution.

The viral particles bind to neurons, enter inside, and release their genetic material into the nucleus, enabling targeted gene expression.

Lidocaine temporarily blocks nerve signals, causing short-term leg paralysis, confirming successful delivery in the CSF.

This protocol establishes intrathecal delivery as an effective method for treating CNS diseases.

Before beginning the procedure, attach a 27-gauge needle to a 25-microliter Hamilton syringe and align the beveled tip of the needle with the volumetric scale on the syringe, then carefully load 8 microliters of 4 times 10 to the 10th gen