We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
siRNA Produced by Recombinant Dicer Mediates Efficient Gene Silencing in Islet Cells.
- Authors
HÄGERKVIST, ROBERT; MOKHTARI, DARIUSH; MYERS, JASON W.; TENGHOLM, ANDERS; WELSH, NILS
- Abstract
RNA interference (RNAi) is emerging as a powerful and convenient tool for studying gene function and genetic variation. RNAi is mediated by 21- to 23-nucleotide-long, small interfering RNAs (siRNA) produced from larger double-stranded RNAs in vivo by the RNase III family enzyme Dicer. To overcome the problems associated with the use of predesigned synthetic siRNA molecules, a novel method utilizing the in vitro activity of recombinant Dicer has been developed recently. In nonislet cells, it has been demonstrated that a pool of siRNA, generated by Dicer from in vitro transcribed dsRNA (d-siRNA), mediates convenient, efficient, and reproducible gene silencing in various cell types. The aim of this study was to evaluate the ability of d-siRNA to silence endogenous gene expression in pancreatic islet cells. We observed that liposomal transfection mediates efficient transport of siRNA in up to 90% of dispersed islet cells and that d-siRNA mediates almost complete and nontoxic silencing of an endogenous mRNA, the messenger coding for the nonreceptor tyrosine kinase c-Abl. The approach described here using d-siRNA provides an important tool for elucidating gene function in further studies of pancreatic islets and diabetes pathophysiology.
- Subjects
HUMAN genetic variation; NUCLEOTIDE sequence; RNA; PROTEIN-tyrosine kinases; GENE expression
- Publication
Annals of the New York Academy of Sciences, 2005, Vol 1040, Issue 1, p114
- ISSN
0077-8923
- Publication type
Article
- DOI
10.1196/annals.1327.014