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BibTeX
Lipid-mediated siRNA delivery down-regulates exogenous gene expression in the mouse brain at picomolar levels
The journal of gene medicine, 2005-02, Vol.7 (2), p.198-207
Hassani, Zahra
Lemkine, Gregory F.
Erbacher, Patrick
Palmier, Karima
Alfama, Gladys
Giovannangeli, Carine
Behr, Jean-Paul
Demeneix, Barbara A.
2005
Details
Autor(en) / Beteiligte
Hassani, Zahra
Lemkine, Gregory F.
Erbacher, Patrick
Palmier, Karima
Alfama, Gladys
Giovannangeli, Carine
Behr, Jean-Paul
Demeneix, Barbara A.
Titel
Lipid-mediated siRNA delivery down-regulates exogenous gene expression in the mouse brain at picomolar levels
Ist Teil von
The journal of gene medicine, 2005-02, Vol.7 (2), p.198-207
Ort / Verlag
Chichester, UK: John Wiley & Sons, Ltd
Erscheinungsjahr
2005
Link zum Volltext
Quelle
Wiley Online Library Journals Frontfile Complete
Beschreibungen/Notizen
Background Efficient in vivo vectors are needed to exploit the enormous potential of RNA interference (RNAi). Such methods require optimisation for specific delivery routes, tissues and usages. We tested the capacity of different non‐viral vectors and formulation methods for inhibition of exogenous (luciferase) gene expression when used to introduce small interfering RNA (siRNA) into the mouse brain in vivo. Methods Polyethylenimine (PEI)‐based polyplexes and JetSI™ (a mixture of cationic lipids)‐based lipoplexes were used to vectorise plasmid DNA encoding the firefly Photinus pyralis luciferase gene and picomolar amounts of siRNA directed against this gene. Two controls were used, DNA encoding an unrelated luciferase from Renilla reniformis and a mutated siRNA sequence. Results First, we found that linear PEI, although efficient for delivering nucleic acids to cells, did not permit development of siRNA activity within the dose range tested (<0.5 pmol). Second, various combinations of cationic lipids were tried and the best formulation was found to be a combination of JetSI™ with the fusogenic lipid dioleoylphosphatidylethanolamine (DOPE). Efficient inhibition of target, firefly luciferase was obtained with exceedingly low amounts of siRNA: 78 ± 6% inhibition at 24 h post‐transfection with 0.2 pmol siRNA. This inhibition was dose‐dependent and specific. No effect was seen on the control gene, co‐transfected Renilla luciferase, and the control mutated siRNA sequence had no effect on the targeted firefly luciferase. Conclusions We have optimised an efficient cationic lipoplex method for delivery of siRNA into the newborn mouse brain. Specific inhibition of exogenous target gene expression is obtained with picomolar amounts of siRNA. Copyright © 2004 John Wiley & Sons, Ltd.
Sprache
Englisch
Identifikatoren
ISSN: 1099-498X
eISSN: 1521-2254
DOI: 10.1002/jgm.659
Titel-ID: cdi_hal_primary_oai_HAL_hal_00181088v1
Format
–
Schlagworte
Analysis of Variance
,
Animals
,
Animals, Newborn
,
Base Sequence
,
Biochemistry, Molecular Biology
,
Brain
,
Brain - metabolism
,
brain, mouse
,
cationic lipids
,
Female
,
Fireflies
,
Fireflies - enzymology
,
Gene Expression Regulation, Enzymologic
,
Gene Expression Regulation, Enzymologic - drug effects
,
Gene Transfer Techniques
,
Genetic Vectors
,
Genetic Vectors - metabolism
,
in vivo
,
Life Sciences
,
Lipid Metabolism
,
Luciferases
,
Luciferases - metabolism
,
Male
,
Mice
,
mouse
,
Particle Size
,
Phosphatidylethanolamines
,
Phosphatidylethanolamines - metabolism
,
Polyethyleneimine
,
Polyethyleneimine - metabolism
,
polyethylenimine
,
Renilla
,
Renilla - enzymology
,
RNA Interference
,
RNA, Small Interfering
,
RNA, Small Interfering - genetics
,
RNA, Small Interfering - metabolism
,
RNA, Small Interfering - pharmacology
,
siRNA
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