This article describes the hydrodynamic gene transfer technique using a mouse model. The method involves rapid intravenous injection of a plasmid solution to facilitate gene delivery to liver cells.
To perform the hydrodynamic gene transfer, begin by taking a syringe filled with a saline-based solution containing recombinant plasmids carrying the gene of interest. Subsequently, take a mouse model and rapidly inject a large volume of plasmid solution at the base of the tail, ensuring the plasmid solution is injected intravenously in the tail vein.
Pressurized injection of a high volume of plasmids generates a mechanical force called the hydrodynamic force, which increases the intravascular pressure in the inferior vena cava - a vein that carries deoxygenated blood to the heart.
Large liquid volume induces congestion in the heart's right ventricle forcing the reversal in blood direction. This step forces the plasmid solution to enter the hepatic vein, which carries it to the liver.
The plasmids move inside the hepatic sinusoids - the spaces in the liver lined by the endothelial cells. Increased blood volume helps the plasmids permeabilize the endothelial layer helping them reach near the surrounding hepatocytes - the liver cells.
The rapid inflow of plasmid solution induces pressure on the hepatocyte cell membrane generating transient pores. These pores facilitate the cells' uptake of the plasmid DNA. Later, the membrane pores close, entrapping the plasmids within the hepatocytes.
The successful expression of the gene of interest in hepatocytes confirms the transfection.
Begin by weighing the mice, then, prepare a sterile saline solution containing 15 micrograms per milliliter of saline endotoxin-free sleeping beauty vector construct and 1 microgram per milliliter endotoxin-free pc-HSB5. Prepare enough solution to inject each mouse with a volume that is 10% of their body weight. Fill a sterile 3-milliliter syringe with the required volume for one mouse, and attach a 27-gauge needle.
After adding an appropriate amount of tissue paper to leave minimal space for movement, but enough space for breathing, insert the mouse into an appropriate restrainer with breathing holes. Warm the tail using an infrared lamp for 30 to 60 seconds, while watching carefully for signs of overheating, such as rapid movement of the tail or other signs of agitation, then, clean the tail with an alcohol swab.
Insert the needle almost horizontally into either one of the two lateral tail veins close to the base of the tail. If placed successfully, a small amount of blood might flow back into the cone of the needle. Inject the total volume into the tail vein within 8 to 10 seconds. After injection, immediately remove the mouse from the restrainer. Compress injection wound with sterile gauze for at least 30 seconds or until any bleeding subsides.
Successful hydrodynamic tail vein injection is crucial for transfection, and especially dependent on injection speed, volume, and time. To make intravenous injection easier, make sure the mice are not stressed, or dehydrated on the day of injection.
After 30 to 60 minutes of solo housing in a recovery cage, transfer the mouse back to its original cage.
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