Hydrodynamic Renal Pelvis Injection: A Technique to Directly Deliver DNA into Kidney Cells of Murine Model

Abstract

Source: Woodard, L. E. et al. Hydrodynamic Renal Pelvis Injection for Non-viral Expression of Proteins in the Kidney. J. Vis. Exp. (2018)

In this video, we present the protocol for hydrodynamic injection of plasmids into the renal pelvis of the mouse kidney for nucleic acid transfection. This technique allows organ-specific delivery of plasmids by utilizing the principle of high hydrostatic pressure, which forcefully drives the plasmids into the specific site without causing tissue injury.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Perform the Hydrodynamic Renal Pelvis Injection Surgery Select mice carefully for surgery. NOTE: Strain-specific differences have not yet been observed but may be possible. Most injections have been on the C57BL/6 or FVB backgrounds. Renal pelvis injections work best in mice that are 6–12 w…

Representative Results

Figure 1. Correct incision site and needle placement for hydrodynamic renal pelvis injections. A) The incision (red line) should be located approximately 1 cm from the spine and approximately 1 cm below the ribcage of the mouse. B) After the kidney is exposed via the flank incision, the renal pelvis should be located as a small yellowish clear/white dot midway down the kidney. Th…

Materials

AnaSed Xylazine	Patterson Veterinary	07-808-1947	Anesthetic – Not controlled substance
BD Insulin Syringe 0.5 mL 29G 1/2 Inch	Cardinal Health	309306	Required syringes
Buprenex	Pharmacist/Veterinarian		Analgesia – Controlled Substance
Extra Fine Micro Dissecting Scissors	Roboz Surgical Instrument	RS-5882	Surgical tool
Gaymar Heat Pump	Paragon Medical	TP-700	Water-circulating heat pump
Germinator 500	Roboz Surgical Instrument	DS-401	To reuse surgical tools during surgery
Graefe Forceps	Roboz Surgical Instrument	RS-5136	Surgical tool
Graefe Tissue Forceps	Roboz Surgical Instrument	RS-5153	Surgical tool
Halsey Needle Holder, 5" Length	Roboz Surgical Instrument	RS-7841	Surgical tool
Heat pads – 15" x 21" – need at least 3	Paragon Medical	TP22G	For use with Gaymar Heat Pump
Ketamine	Pharmacist/Veterinarian		Anesthetic – Controlled Substance
Prevantics Swabs	Thermo Fisher Scientific	23-100-110	For skin surgery prep
Prolene 5-0 sutures Taper 30"	Thermo Fisher Scientific	NC0256891	Non-absorbable sutures for skin
Puralube Brand Opthalmic Ointment	Patterson Veterinary	07-888-2572	To keep eyes moist during surgery
Vicryl 5-0 Sutures J303H	Thermo Fisher Scientific	NC9816710	Absorbable sutures for muscle layer
Wahl Mini Arco Clipper	Med-Vet Internationa	I 8787-1550	Shaver for skin prep