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

Abstract

Source: Fricano-Kugler, C. J., et al. Designing, Packaging, and Delivery of High Titer CRISPR Retro and Lentiviruses via Stereotaxic Injection. J. Vis. Exp. (2016)

This video demonstrates stereotaxic viral injection for precise neuronal gene editing in the mouse brain. The protocol ensures accurate virus delivery to the dentate gyrus, enabling fluorescent labeling and CRISPR-mediated gene knockout to study neural circuits, gene function, and disease mechanisms in vivo.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Stereotaxic Injection

1. Confirm that the ventilation to the surgery suite is open to ensure proper airflow. Prepare the mouse for surgery by anesthetizing it with 4% isoflurane in an induction chamber. Following anesthesia, shave the head to prepare the injection site.
   1. Place the mouse in the stereotaxic instrument by using tweezers to move the tongue down and to the side. Insert the bite bar into the mouth until the teeth drop into the slot, then secure the ear bars. Ensure that the body o....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Stereotaxic Surgery Reagents
Vet-Syringe vet use T.B. Syringe only 1cc luer slip T.B. 100/bx	Med-Vet International	1CCVLS
Isoflurane
Stainless Steel Scalpel Blades, #10, 100-pk	Med-Vet International	JOR580S
Artificial tear ointment	Med-Vet International	RXPARALUBE-O
PVP PrepSolution	Med-Vet International	HPIV108208H
Normal saline	Med-Vet International	DYND500MLSH
Cotton tipped applicators	Med-Vet International	CTA6
Triple antibiotic ointment	Med-Vet International	RXTRIP-OI15
MONOJECT® Needles Soft Pack 25g x 5/8"	Med-Vet International	25058
6-0 silk sutures	Med-Vet International	MV-711