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. 2010 Dec;16(12):2435–2441. doi: 10.1261/rna.2347310

FIGURE 1.

FIGURE 1.

Inhibitors of a trypanosomatid editing reaction. (A) The RNA reporter for the HTS contains a streptavidin aptamer (yellow) that is only active after the insertion of three U's (red) by the in vitro editing reaction. Upon immobilization of the RNA at the bottom of a streptavidin-coated microtiter plate and electrical stimulation, an ECL signal is generated from a ruthenium complex attached to the reporter. (B) Confirmation of the inhibitory activity using a radiolabeled reporter that is based on the cytochrome b mRNA sequence. The gRNA used for the assay has two guiding nucleotides. Compounds were dissolved in DMSO and used at a final concentration of 10 μM. Reactions contained 0.01% TX-100, and products were analyzed after electrophoresis on a denaturing gel. No editing activity was detected in the presence of any of the inhibitors (n = 3). (C) Confirmation of the inhibitory activity of compounds obtained from the HTS using a radiolabeled SELEX-optimized reporter with two guiding nucleotides. Reactions contained 0.01% TX-100, and products were analyzed after electrophoresis on a denaturing gel. The percent inhibition for correct editing (+2) in the presence of each compound is relative to the DMSO-only control (n = 3).