A method to facilitate and monitor expression of exogenous genes in the rat kidney using plasmid and viral vectors

Abstract

Corridon PR, Rhodes GJ, Leonard EC, Basile DP, Gattone VH II, Bacallao RL, Atkinson SJ. A method to facilitate and monitor expression of exogenous genes in the rat kidney using plasmid and viral vectors.

Kidney-selective gene transfection using anionic bubble lipopolyplexes with renal ultrasound irradiation in mice

“…10,[28][29][30] Proximal tubule epithelial cells were reported to have a huge capacity for apical endocytic uptake of exogenous materials, but most gene delivery carriers could not pass through the glomerulus filtration barrier. 31 Therefore, genes can be transferred into the tubules not only by injection into the renal artery but by injection into the renal parenchyma and retrograde injection from the ureter. 8,9,32 Antisense oligonucleotides can be successfully delivered to the kidney vasculature using cationic liposomes, 33 and genes have delivered to the renal interstitium hydrodynamically by retrograde renal vein injection.…”

Hydrodynamic Renal Pelvis Injection for Non-viral Expression of Proteins in the Kidney

“…Future electron microscopy studies may elucidate this mechanism further. The retrograde nature of the injection has precedence in that the hydrodynamic tail vein injection method reverses the flow of the hepatic vein 20 and other successful methods of kidney transfection have injected via the renal vein 12–14,21 . Kidney damage is likely required for optimal transfection efficiencies so we hypothesize that transfection occurs via damage to the cell membrane caused by the fast, high-volume injection.…”

“…Additionally, both the piggyBac transposon system and/or cyclophosphamide treatment to suppress the immune reaction to the transgene have been shown to improve long-term gene expression outcomes 11 . Other investigators have used a renal vein approach in rat with success, achieving high transfection efficiency for time periods of greater than one month 14 . However, genetic correction of phenotypes mimicking human disease are usually performed in mice first as a proof-of-concept since most mammalian genetic models are mouse models.…”

Intravital imaging of the kidney

“…Understanding renal physiology and using proper controls are crucial to establish a reproducible experimental system. We did not discuss the use of plasmids or viruses as a means to deliver probes to the kidney because of the lack of established methods to date [10,35–40]. A simple injection of plasmids (via tail vein or jugular vein) is not effective due to trapping by other organs.…”