@article {825736, title = {AutomiG, a biosensor to detect alterations in miRNA biogenesis and in small RNA silencing guided by perfect target complementarity.}, journal = {PLoS One}, volume = {8}, year = {2013}, month = {2013}, pages = {e74296}, 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.

}, keywords = {Animals, Argonaute Proteins, Biosensing Techniques, Cell Line, Drosophila, Drosophila Proteins, Green Fluorescent Proteins, MicroRNAs, Promoter Regions, Genetic, RNA Interference}, issn = {1932-6203}, doi = {10.1371/journal.pone.0074296}, author = {Carr{\'e}, Cl{\'e}ment and Jacquier, Caroline and Boug{\'e}, Anne-Laure and de Chaumont, Fabrice and Besnard-Guerin, Corinne and Thomassin, H{\'e}l{\`e}ne and Pidoux, Josette and Da Silva, Bruno and Chalatsi, Eleftheria and Zahra, Sarah and Olivo-Marin, Jean-Christophe and Munier-Lehmann, H{\'e}l{\`e}ne and Antoniewski, Christophe} }