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

J Vis Exp. 2018 Jan 8;(131):56324. doi: 10.3791/56324.

Abstract

Hydrodynamic injection creates a local, high-pressure environment to transfect various tissues with plasmid DNA and other substances. Hydrodynamic tail vein injection, for example, is a well-established method by which the liver can be transfected. This manuscript describes an application of hydrodynamic principles by injection of the mouse kidney directly with plasmid DNA for kidney-specific gene expression. Mice are anesthetized and the kidney is exposed by a flank incision followed by a fast injection of a plasmid DNA-containing solution directly into the renal pelvis. The needle is kept in place for ten seconds and the incision site is sutured. The following day, live animal imaging, Western blot, or immunohistochemistry may be used to assay gene expression, or other assays suited to the transgene of choice are used for detection of the protein of interest. Published methods to prolong gene expression include transposon-mediated transgene integration and cyclophosphamide treatment to inhibit the immune response to the transgene.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Video-Audio Media

MeSH terms

  • Animals
  • DNA / administration & dosage*
  • DNA / genetics
  • Hydrodynamics
  • Injections
  • Kidney / metabolism
  • Kidney / physiology*
  • Kidney Pelvis / physiology*
  • Male
  • Mice
  • Plasmids / administration & dosage*
  • Plasmids / genetics
  • Protein Biosynthesis / genetics*
  • Transfection / methods*
  • Transgenes

Substances

  • DNA