Absence of transitive and systemic pathways allows cell-specific and isoform-specific RNAi in Drosophila.

Details

Serval ID
serval:BIB_DF5A64864AE5
Type
Article: article from journal or magazin.
Collection
Publications
Title
Absence of transitive and systemic pathways allows cell-specific and isoform-specific RNAi in Drosophila.
Journal
RNA
Author(s)
Roignant J.Y., Carré C., Mugat B., Szymczak D., Lepesant J.A., Antoniewski C.
ISSN
1355-8382 (Print)
ISSN-L
1355-8382
Publication state
Published
Issued date
03/2003
Peer-reviewed
Oui
Volume
9
Number
3
Pages
299-308
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Abstract
RNA interference (RNAi) designates the multistep process by which double-stranded RNA induces the silencing of homologous endogenous genes. Some aspects of RNAi appear to be conserved throughout evolution, including the processing of trigger dsRNAs into small 21-23-bp siRNAs and their use to guide the degradation of complementary mRNAs. Two remarkable features of RNAi were uncovered in plants and Caenorhabditid elegans. First, RNA-dependent RNA polymerase activities allow the synthesis of siRNA complementary to sequences upstream of or downstream from the initial trigger region in the target mRNA, leading to a transitive RNAi with sequences that had not been initially targeted. Secondly, systemic RNAi may cause the targeting of gene silencing in one tissue to spread to other tissues. Using transgenes expressing dsRNA, we investigated whether transitive and systemic RNAi occur in Drosophila. DsRNA-producing transgenes targeted RNAi to specific regions of alternative mRNA species of one gene without transitive effect directed to sequences downstream from or upstream of the initial trigger region. Moreover, specific expression of a dsRNA, using either cell-specific GAL4 drivers or random clonal activation of a GAL4 driver, mediated a cell-autonomous RNAi. Together, our results provide evidence that transitive and systemic aspects of RNAi are not conserved in Drosophila and demonstrate that dsRNA-producing transgenes allow powerful reverse genetic approaches to be conducted in this model organism, by knocking down gene functions at the resolution of a single-cell type and of a single isoform.
Keywords
Animals, Animals, Genetically Modified, DNA Probes, Drosophila/genetics, Drosophila Proteins/genetics, Gene Expression Regulation, Green Fluorescent Proteins, Luminescent Proteins/genetics, Microscopy, Confocal, Protein Isoforms, RNA Interference, Receptors, Steroid/genetics, Repetitive Sequences, Nucleic Acid, Transcription Factors/genetics, Transgenes
Pubmed
Web of science
Create date
28/10/2019 13:02
Last modification date
29/10/2019 6:26
Usage data