Other Publications (1)
Articles by Felisha M. Williams in JoVE
Hydrodynamic Renal Pelvis Injection for Non-viral Expression of Proteins in the Kidney Lauren E. Woodard1,2,3, Richard C. Welch2, Felisha M. Williams2, Wentian Luo2, Jizhong Cheng3, Matthew H. Wilson1,2,3 1Department of Veterans Affairs, Tennessee Valley Healthcare System, 2Departments of Medicine and Pharmacology, Vanderbilt University Medical Center, 3Department of Medicine, Baylor University College of Medicine This protocol describes a method to inject plasmid DNA into the mouse kidney via the renal pelvis to produce transgene expression specifically in the kidney.
Other articles by Felisha M. Williams on PubMed
Kidney-specific Transposon-mediated Gene Transfer in Vivo Scientific Reports. | Pubmed ID: 28317878 Methods enabling kidney-specific gene transfer in adult mice are needed to develop new therapies for kidney disease. We attempted kidney-specific gene transfer following hydrodynamic tail vein injection using the kidney-specific podocin and gamma-glutamyl transferase promoters, but found expression primarily in the liver. In order to achieve kidney-specific transgene expression, we tested direct hydrodynamic injection of a DNA solution into the renal pelvis and found that luciferase expression was strong in the kidney and absent from extra-renal tissues. We observed heterogeneous, low-level transfection of the collecting duct, proximal tubule, distal tubule, interstitial cells, and rarely glomerular cells following injection. To assess renal injury, we performed the renal pelvis injections on uninephrectomised mice and found that their blood urea nitrogen was elevated at two days post-transfer but resolved within two weeks. Although luciferase expression quickly decreased following renal pelvis injection, the use of the piggyBac transposon system improved long-term expression. Immunosuppression with cyclophosphamide stabilised luciferase expression, suggesting immune clearance of the transfected cells occurs in immunocompetent animals. Injection of a transposon expressing erythropoietin raised the haematocrit, indicating that the developed injection technique can elicit a biologic effect in vivo. Hydrodynamic renal pelvis injection enables transposon mediated-kidney specific gene transfer in adult mice.