Recombinant Adeno Associated Virus Based Gene Delivery: A Technique for Transgene Delivery and Cardiac-Specific Expression in Rodent Model
Published: April 30, 2023
Abstract
Source: Ding, J. et al. Preparation of rAAV9 to Overexpress or Knockdown Genes in Mouse Hearts. J. Vis. Exp. (2016)
In this video, we demonstrate recombinant adeno associated virus-based gene delivery in the mouse pup. This method helps in studying gene expression in the targeted cells.
Protocol
1. Treat neonatal mice with rAAV9 at P0.5-P2.5 by subcutaneous injection.
- Pre-fill a 29G1/2, 0.33 x 12.7 mm insulin syringe with the recombinant adeno-associated virus 9 (rAAV9) solution. Be careful to remove air bubbles.
- Hold the cryo-anesthetized pup in one hand with thumb and forefinger. Prior to injection, swipe the back skin of the pup with a swab stick saturated with 70% isopropyl alcohol to maintain the sterile condition. Insert the syringe needle into the anterior-dorsal subcutis of the animal at an angle of 5 to 10°. Inject 50-70 µl of the rAAV9 solution using the insulin syringe.
NOTE: rAAV9 can also be delivered to the mouse via intraperitoneal or intravenous injection. Efficient expression of the delivered genes in the heart can be obtained. However, intraperitoneal injection sometimes may result in leaky expression in the liver. After injection, the condition of the pups was monitored every day.
Divulgaciones
The authors have nothing to disclose.
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Recombinant Adeno Associated Virus Based Gene Delivery: A Technique for Transgene Delivery and Cardiac-Specific Expression in Rodent Model. J. Vis. Exp. (Pending Publication), e20986, doi: (2023).