Stereotaxic Injection of Viral Vectors for Glial Cell Reprogramming in a Mouse Model

Secure an anesthetized transgenic mouse in a stereotaxic frame with its skull exposed.

This mouse expresses Cre recombinase in glial cells.

Position the stereotaxic arm holding the capillary tube over the skull and align it above the bregma.

Adjust the stereotactic axes for precise targeting.

Drill a hole at the target site, ensuring the dura mater remains intact. Remove bone fragments.

Flush the capillary tube with buffer. Draw up an air bubble, then load the adeno-associated virus carrying inactive neuronal reprogramming and fluorescent protein genes.

Lower the capillary tube to the target depth and inject the viral solution into the glial cell-rich region.

Carefully retract the tube to prevent backflow.

Close the incision and allow recovery.

The internalized viruses deliver loxP-flanked inactivated genes to the glial nucleus. Cre recombinase flips them, activating neuronal reprogramming genes and driving neuronal conversion. Reprogrammed interneurons express the fluorescent protein under the neuron-specific promoter.

To inject reprogramming factors, place an anesthetized mouse in the stereotaxic frame, administer appropriate analgesia at the beginning of the surgery, then bring the tip o