In Utero Viral Transduction in a Mouse

Take a mixture of virus containing a desired gene for transduction and a tracking dye in a buffer.

Aspirate the virus-dye mixture into a fine-tip capillary.

Take an anesthetized pregnant mouse with a shaved abdominal region.

Disinfect the skin with ethanol, followed by a betadine solution.

Make a midline incision to expose the abdominal cavity, and place a paraffin film over it.

Expose the uterine horns and pour buffer over them to prevent dryness.

Note the order and position of the embryos. Adjust the embryo's position within the uterine sac to access the injection sites.

Inject the virus-dye mixture into the spinal cord and brain ventricles.

Ensure accurate injection by visualizing the dye distribution.

Place the uterine horns back into the abdominal cavity and pour some buffer.

Suture the abdominal cavity, followed by the skin.

The injected virus enters the host cell, releasing the target gene and enabling its expression.

Begin by loading a customized capillary tip containing four times 10 to the 11 viral particles of adeno-associated virus type 1 coating for the desired target, and labeled with Fast Green dye onto an aspirator tube. Add 15 microliters of the labeled adeno-associated virus