ARS | Publication request: Characterizing siRNA production from a dual reporter system

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: October 23, 2006
Publication Date: January 18, 2007
Citation: Chapa, D.L., Velten, J.P. 2007. Characterizing siRNA production from a dual reporter system [abstract]. Plant Biology 2007. January 18-20, 2007, Riverside, California. CD-ROM.

Technical Abstract: Various reporter genes have proven effective at demonstrating the effects of RNA silencing in plants. Previous data has indicated a differential effect of RNA silencing on two luciferase genes (unrelated at the DNA sequence level), irrespective of the promoters use to drive the reporter genes. To investigate the observed preferential silencing, labeled siRNAs isolated from wild type N. benthamiana leaf tissue (infused with A. tumefaciens harboring T-DNA containing a 35S::Firefly luciferase and SuperPromoter::Renilla luciferase) were analyzed by Slot Blot hybridization to DNA segments of the full Luciferase transcripts. The purpose of this research is to: 1) reveal the segments of the luciferase transcripts that are targeted by the different siRNAs; and 2) correlate the hybridization signal strength and distribution to silencing previously measured at the mRNA and protein levels. Quantification of any bias as to how siRNAs target the foreign nucleic acid will enhance our understanding of the mechanisms behind siRNA production and target specificity.