Skip to Main content Skip to Navigation
Journal articles

AutomiG, a biosensor to detect alterations in miRNA biogenesis and in small RNA silencing guided by perfect target complementarity.

Abstract : Defects in miRNA biogenesis or activity are associated to development abnormalities and diseases. In Drosophila, miRNAs are predominantly loaded in Argonaute-1, which they guide for silencing of target RNAs. The miRNA pathway overlaps the RNAi pathway in this organism, as miRNAs may also associate with Argonaute-2, the mediator of RNAi. We set up a gene construct in which a single inducible promoter directs the expression of the GFP protein as well as two miRNAs perfectly matching the GFP sequences. We show that self-silencing of the resulting automiG gene requires Drosha, Pasha, Dicer-1, Dicer-2 and Argonaute-2 loaded with the anti-GFP miRNAs. In contrast, self-silencing of the automiG gene does not involve Argonaute-1. Thus, automiG reports in vivo for both miRNA biogenesis and Ago-2 mediated silencing, providing a powerful biosensor to identify situations where miRNA or siRNA pathways are impaired. As a proof of concept, we used automiG as a biosensor to screen a chemical library and identified 29 molecules that strongly inhibit miRNA silencing, out of which 5 also inhibit RNAi triggered by long double-stranded RNA. Finally, the automiG sensor is also self-silenced by the anti-GFP miRNAs in HeLa cells and might be easily used to identify factors involved in miRNA biogenesis and silencing guided by perfect target complementarity in mammals.
Document type :
Journal articles
Complete list of metadatas

Cited literature [48 references]  Display  Hide  Download

https://hal-pasteur.archives-ouvertes.fr/pasteur-00877254
Contributor : Hélène Munier-Lehmann <>
Submitted on : Monday, October 28, 2013 - 11:58:43 AM
Last modification on : Monday, August 31, 2020 - 12:36:19 PM
Long-term archiving on: : Friday, April 7, 2017 - 5:43:48 PM

File

journal.pone.0074296.pdf
Publisher files allowed on an open archive

Identifiers

Citation

Clément Carré, Caroline Jacquier, Anne-Laure Bougé, Fabrice de Chaumont, Corinne Besnard-Guerin, et al.. AutomiG, a biosensor to detect alterations in miRNA biogenesis and in small RNA silencing guided by perfect target complementarity.. PLoS ONE, Public Library of Science, 2013, 8 (9), pp.e74296. ⟨10.1371/journal.pone.0074296⟩. ⟨pasteur-00877254⟩

Share

Metrics

Record views

1024

Files downloads

1431