Hydrodynamic Renal Pelvis Injection: A Technique to Directly Deliver DNA into Kidney Cells of Murine Model
Transcription
To deliver plasmid DNA directly into the mouse kidney cells, begin by placing an anesthetized mouse in a lateral position. Prep the mouse by removing its dorsal fur from the side. Locate an appropriate incision site in the left flank, below the ribcage.
Make a small slit in the skin and underlying muscle layers. Next, apply gentle pressure on the abdominal cavity to expose the kidney through the incision site. Partially remove the overlaying perinephric fat tissue to visualize the renal pelvis.
Now, take a syringe containing plasmid DNA in a hydrodynamic delivery buffer. Insert the needle directly into the renal pelvis – a triangular, dilated portion of the ureter in the kidney. Inject the plasmid solution rapidly such that the DNA directly reaches the renal parenchyma – the functional part of the kidney that filters blood.
The quick delivery of large fluid volume through the renal pelvis builds a high pressure, creating transient pores in the plasma membrane of renal parenchymal cells. This facilitates the entry of plasmids into the cells. Eventually, these pores close, entrapping the plasmids within the cells.
Finally, retract the needle and return the kidney to the abdominal cavity. Suture the skin and muscle layers at the incision site.
