Highly Efficient Small Interfering RNA Delivery to Primary Mammalian Neurons Induces MicroRNA-Like Effects before mRNA Degradation

The study of protein function in neurons has been hindered by the lack of highly efficient, nontoxic methods of inducing RNA interference in such cells. Here we show that application of synthetic small interfering RNA (siRNA) linked to the vector peptide Penetratin1 results in rapid, highly efficien...

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Published inThe Journal of neuroscience Vol. 24; no. 45; pp. 10040 - 10046
Main Authors Davidson, Thomas J, Harel, Sivan, Arboleda, Valerie A, Prunell, Giselle F, Shelanski, Michael L, Greene, Lloyd A, Troy, Carol M
Format Journal Article
LanguageEnglish
Published United States Soc Neuroscience 10.11.2004
Society for Neuroscience
Subjects
Animals
Carrier Proteins - administration & dosage
Caspase 3
Caspases - analysis
Caspases - genetics
Cells, Cultured - drug effects
Cells, Cultured - metabolism
Cellular/Molecular
Culture Media, Serum-Free - pharmacology
Gene Targeting - methods
Hippocampus - cytology
Mice
Neurons - drug effects
Neurons - metabolism
Protein Biosynthesis - drug effects
Rats
RNA, Messenger - metabolism
RNA, Small Interfering - pharmacology
Superior Cervical Ganglion - cytology
Superoxide Dismutase - analysis
Superoxide Dismutase - genetics
Superoxide Dismutase-1
Transfection
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Summary:The study of protein function in neurons has been hindered by the lack of highly efficient, nontoxic methods of inducing RNA interference in such cells. Here we show that application of synthetic small interfering RNA (siRNA) linked to the vector peptide Penetratin1 results in rapid, highly efficient uptake of siRNA by entire populations of cultured primary mammalian hippocampal and sympathetic neurons. This treatment leads to specific knock-down of targeted proteins within hours without the toxicity associated with transfection. In contrast to current methods, our technique permits study of protein function across entire populations with minimal disturbance of complex cellular networks. Using this technique, we found that protein knock-down (evident after 6 hr) precedes any decrease in targeted message (evident after 24 hr), suggesting an early, translational repression by perfectly targeted siRNAs.
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ISSN:0270-6474
1529-2401
1529-2401
DOI:10.1523/JNEUROSCI.3643-04.2004