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. 2008 May 23;9(8):1225-8.
doi: 10.1002/cbic.200800051.

Light-regulated RNA-small molecule interactions

Douglas D Young  1 , Alexander Deiters
Affiliations

Light-regulated RNA-small molecule interactions

Douglas D Young et al. Chembiochem. .
No abstract available

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Figures

Figure 1
Figure 1
Specificity assay for isolated aptamers. Isotopically labeled RNA was incubated with the spiropyran resin 1, then washed, resuspended in fresh buffer, and radioactivity in the supernatant was quantitated (dark bars), the resin was then switched to 2a through irradiation at 365 nm, and the supernatant was quantitated again (light bars). The RNA aptamer SP3 demonstrated the greatest specificity for spiropyran 1 over the merocyanine 2a. RNA=aptamer concentration.
Figure 2
Figure 2
Surface plasmon resonance experiment demonstrating the reversibility of the light-regulated SP3 aptamer binding. The spiropyran 1 was immobilized on a gold surface, incubated with the SP3 aptamer, and irradiated at 365 nm. SPR=change in percent reflectivity.
Scheme 1
Scheme 1
Spiropyran 1 (colorless) and its merocyanine forms 2a (purple) and 2b (yellow).
Scheme 2
Scheme 2
Photochemical in vitro selection. An RNA library is incubated with a resin containing the spiropyran 1 (square). RNA aptamers incapable of binding are washed away and the resin is switched to the merocyanine form 2a (circle) with UV light of 365 nm. RNAs which are nonspecifically bound, or recognize both isomers are retained, whereas specific binders are eluted and collected. This enriched pool of RNA aptamers is then reverse transcribed, PCR amplified, and transcribed back to RNA to continue the cycle. The selection cycle was performed ten times until significant enrichment of the RNA pool was detected.
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