Stereotaxic Viral Injection for Targeted Neuronal Gene Editing in the Mouse Brain

Secure an anesthetized mouse in a stereotaxic frame.

Disinfect the shaved scalp, then make a midline incision to expose the skull.

Dry the skull and apply hydrogen peroxide to enhance bregma visibility.

Locate bregma and lambda, then identify the target sites.

Carefully drill through the skull at the target sites.

Attach the virus-filled syringe to the stereotaxic instrument.

The virus encodes Cas9 and a single-guide RNA for genome editing in dentate gyrus neurons, along with a fluorescent reporter to label infected neurons.

Lower the syringe to reach the dentate gyrus, then inject the virus slowly to ensure uniform distribution.

Gradually raise the syringe and repeat the injection at additional coordinates.

Pause briefly to prevent backflow, then withdraw the needle slowly.

Repeat the procedure on the opposite hemisphere.

Finally, suture the incision.

The model is now ready for further studies.

Shave the head of an anesthetized mouse in order to prepare for the injection site. Direct 4% isoflurane into the nose cone. Next, place the mouse in the stereotaxic instrument. Insert the bite bar into its mouth until its teeth drop into the slot before securing the ear bars.

Ensure that the animal's b