Lee, K. H., Kimoto, M., Kawai, G., Okamoto, I., Fin, A., & Hirao, I. (2022). Dye‐Conjugated Spinach RNA by Genetic Alphabet Expansion. Chemistry – A European Journal. Portico. https://doi.org/10.1002/chem.202104396
Light-emitting systems using an RNA aptamer-dye pair, such as Spinach RNA, are an attractive method for imaging and tracing RNA expression in vitro and in vivo . We present an alternative Spinach method by genetic alphabet expansion using an unnatural base pair system, in which a dye-conjugated unnatural base substrate is site-specifically incorporated at a specific position in Spinach RNA by transcription involving the third base pair. The incorporation position was predicted by molecular dynamics simulations. This dye-conjugated Spinach RNA increased the thermal stability of the fluorescence, the robustness against ion sensitivity, and the resistance against photobleaching. Furthermore, we applied our method to Baby Spinach, a shorter version of Spinach, for dye conjugation toward the visible detection of transcripts. This is the first demonstration of an alternative RNA imaging method for a detection system using genetic alphabet expansion.
This research is supported by core funding from: Institute of Bioengineering and Bioimaging
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This is the peer reviewed version of the following article: Lee, K. H., Kimoto, M., Kawai, G., Okamoto, I., Fin, A., & Hirao, I. (2022). Dye‐Conjugated Spinach RNA by Genetic Alphabet Expansion. Chemistry – A European Journal. Portico. https://doi.org/10.1002/chem.202104396, which has been published in final form at http://dx.doi.org/10.1002/chem.202104396. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions