Hydrodynamic Renal Pelvis Injection: A Technique to Directly Deliver DNA into Kidney Cells of Murine Model
Published: April 30, 2023
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…
Divulgaciones
The authors have nothing to disclose.
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 |
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Hydrodynamic Renal Pelvis Injection: A Technique to Directly Deliver DNA into Kidney Cells of Murine Model. J. Vis. Exp. (Pending Publication), e20569, doi: (2023).