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The Journal of Neuroscience, November 10, 2004, 24(45):10040-10046; doi:10.1523/JNEUROSCI.3643-04.2004

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

Thomas J. Davidson,¹ Sivan Harel,¹ Valerie A. Arboleda,¹ Giselle F. Prunell,¹ Michael L. Shelanski,^1,3 Lloyd A. Greene,^1,3 and Carol M. Troy^1,2,3

Departments of ¹Pathology and ²Neurology, Taub Institute for the Study of Alzheimer's Disease and the Aging Brain, and ³Center for Neurobiology and Behavior, Columbia University College of Physicians and Surgeons, New York, New York, 10032

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.

Key words: primary neurons; hippocampal neurons; siRNA; Penetratin1; transduction peptides; RNA interference

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Received July 19, 2004; accepted September 21, 2004.

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