Hydrodynamic Gene Delivery: A Technique for In Vivo Transfection of Exogenous DNA in Murine Hepatocytes via Tail Vein Injection
Transcription
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.
