Prolonged in vivo gene silencing by electroporation-mediated plasmid delivery of small interfering RNA

Prolonged in vivo gene silencing by electroporation-mediated plasmid delivery of small interfering RNA

Eefting, D.
Grimbergen, J.M.
de Vries, M.R.
van Weel, V.
Kaijzel, E.L.
Que, I.
Moon, R.T.
Löwik, C.W.
van Bockel, J.H.
Quax, P.H.A.
TNO Kwaliteit van Leven

2007

For the successful application of RNA interference in vivo, it is desired to achieve (local) delivery of small interfering RNAs (siRNAs) and long-term gene silencing. Nonviral electrodelivery is suitable to obtain local and prolonged expression of transgenes. By intramuscular electrodelivery of a plasmid in which two opposing human polymerase III promoters (H1 and U6) drive the expression of siRNA constructs that form functional double-stranded siRNAs, in combination with in vivo bioluminescence imaging, we were able to knock down exogenous delivered luciferase for at least 100 days in murine calf muscles. This effect was sequence specific, because scrambled siRNA had no effect. Moreover, we were able to demonstrate in vivo reduction of endogenous TLR4 expression for at least 1 week, using a similar vector expressing an siRNA for TLR4 in the muscle. In this study, we demonstrate that in vivo suppression of both endogenous (for at least 1 week) and introduced genes (>100 days) is feasible via plasmid-driven siRNA expression after electroporation-mediated intramuscular gene transfer. With this approach the short-term effect of oligonucleotides and the drawbacks of viral gene delivery, like immunological responses, could be circumvented. Therefore, this application of RNA interference is a useful tool with which to investigate gene function and might be promising as a therapeutic tool for locally acting diseases such as restenosis or tumors. © Mary Ann Liebert, Inc.

Biology
Biomedical Research
double stranded RNA
histidine
luciferase
oligonucleotide
plasmid vector
small interfering RNA
toll like receptor 4
uridine
animal cell
animal experiment
article
bioluminescence
controlled study
electroporation
gastrocnemius muscle
gene expression
gene function
gene repression
gene silencing
human
human cell
immune response
in vivo gene transfer
in vivo study
male
mouse
nonhuman
nonviral gene delivery system
protein expression
restenosis
RNA interference
transgene
treatment planning
tumor
viral gene delivery system
Animals
Cattle
Cell Line, Transformed
Cell Transformation, Viral
Electroporation
Feasibility Studies
Gene Silencing
Genes, Reporter
Humans
Hypoxanthine Phosphoribosyltransferase
Lipopolysaccharides
Luciferases
Luminescent Measurements
Male
Mice
Mice, Inbred Strains
Muscle, Skeletal
NIH 3T3 Cells
Plasmids
RNA Interference
RNA, Messenger
RNA, Small Interfering
Time Factors
Toll-Like Receptor 4
Murinae

http://resolver.tudelft.nl/uuid:4fa1129a-ff4f-4d73-849f-cfb9f0d2e457

https://doi.org/10.1089/hum.2006.176

240177

1043-0342

Human Gene Therapy, 18 (9), 861-869

article