Stereotaxic Injection of Viruses for Stable Genetic Modification in a Mouse Model

Begin with an anesthetized mouse with an exposed skull in a stereotaxic frame.

Lower the syringe holder until the needle tip aligns with the bregma, a skull reference point.

Using the pre-determined stereotaxic coordinates, locate the ventricular-subventricular zone or

V-SVZ, a neurogenic niche primarily containing neural stem cells or NSCs, and mark this region.

Remove the syringe and carefully drill a small hole, avoiding brain damage.

Fill a sterile syringe with viruses carrying the transgene and position the needle over the brain surface.

Reposition the syringe and lower it until the tip touches the innermost brain surface layer.

Penetrate the brain to reach the V-SVZ, slowly inject the virus, hold briefly, and then withdraw the syringe.

In the V-SVZ, the virus enters NSCs via membrane fusion. Its RNA reverse transcribes into DNA.

This DNA then integrates into the NSC genome, enabling stable genetic modification of NSCs.

After anesthetizing, a six to eight week old mouse shave the area between its ears. Next, disinfect the skin using an iodophor such as iodine povidone or 70% ethanol. Then, place the animal in a prone position on a stereotaxic frame.

Carefully fix its head using the ear b