Specific interference with gene expression and gene function mediated by long dsRNA in neural cells.

TitleSpecific interference with gene expression and gene function mediated by long dsRNA in neural cells.
Publication TypeJournal Article
Year of Publication2002
AuthorsGan L, Anton KE, Masterson BA, Vincent VAM, Ye S, Gonzalez-Zulueta M
JournalJ Neurosci Methods
Date Published2002 Dec 15
KeywordsAnimals, Cell Culture Techniques, Gene Silencing, Green Fluorescent Proteins, Luminescent Proteins, Mice, Microtubule-Associated Proteins, Neuroblastoma, Neurons, Poly(ADP-ribose) Polymerases, RNA Interference, RNA, Double-Stranded, RNA, Small Interfering, Time Factors, Tretinoin, Tumor Cells, Cultured

Double-stranded (ds) RNA-induced sequence-specific interference with gene expression, RNA interference (RNAi), has been extensively used in invertebrates, allowing for efficient and high-throughput gene silencing and gene function analysis. In vertebrates, however, use of RNAi to study gene function has been limited due to non-specific effects induced by double-stranded RNA (dsRNA)-dependent protein kinase and interferon activation. dsRNA-induced specific inhibition of vertebrate gene expression has only been shown in embryonic and non-differentiated mammalian cells. In this report, we demonstrate dsRNA-induced specific interference of gene expression and gene function in partially as well as fully differentiated mouse neuroblastoma cells. Specific silencing was observed in the expression of an integrated transgene coding for green fluorescent protein and a variety of endogenous genes. Moreover, we show that RNAi-mediated inhibition of poly (ADP-ribose) polymerase (PARP) expression induced cellular resistance to oxygen-glucose deprivation, consistent with the role of PARP in ischemia-induced brain damage. Our results indicate that RNAi can be used as a powerful tool to study gene function in neural cells.

Alternate JournalJ. Neurosci. Methods
PubMed ID12468005