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Publication
Kidney-specific transposon-mediated gene transfer in vivo.
Authors Woodard LE, Cheng J, Welch RC, Williams FM, Luo W, Gewin LS, Wilson MH
Submitted By Submitted Externally on 3/30/2017
Status Published
Journal Scientific reports
Year 2017
Date Published 3/1/2017
Volume : Pages 7 : 44904
PubMed Reference 28317878
Abstract 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.




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