Delivery of DNA Plasmids into Urothelium of Mouse Bladder: An Electroporation-based Procedure to Efficiently Deliver DNA Plasmids into Urothelial Cells of Murine Bladder to Generate Genetically Engineered Models

To begin, prep an anesthetized female mouse in the supine position. Ensure its bladder is completely empty. Through the urethral opening, insert a lubricated catheter. Lubrication aids in smooth urethral passage for entry into the bladder.

Following catheterization, make an abdominal incision and dissect the inner musculature to expose the bladder. Next, pipette a suspension of DNA plasmids mixed with a suitable dye into the outer end of the catheter. These DNA plasmids code for a fluorescent reporter gene.

Attach a syringe to the catheter and push it to deliver the plasmid solution into the bladder. The injected dye molecules color the bladder, confirming the successful administration of plasmid solution. Remove the catheter assembly and tighten the external urethral orifice to prevent the plasmid solution out-flow.

Now, pinch the bladder using two electrodes connected to an electroporation generator. Apply electrical pulses of appropriate voltage for a desired period to the bladder. The exposure of the bladder urothelium, the innermost lining of the bladder, to the applied electric fields results in temporary pores in the urothelial cell membrane.

The increased membrane permeability allows the efficient delivery of the injected DNA plasmids into the cells. Following electroporation, suture the surgical incision. Allow the mouse to recover. The urothelial cells transfected with the DNA plasmids express fluorescent reporter proteins.