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Dye‐Conjugated Spinach RNA by Genetic Alphabet Expansion
Author(s) -
Lee Kyung Hyun,
Kimoto Michiko,
Kawai Gota,
Okamoto Itaru,
Fin Andrea,
Hirao Ichiro
Publication year - 2022
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.202104396
Subject(s) - spinach , rna , photobleaching , transcription (linguistics) , conjugated system , dna , chemistry , base pair , biology , biophysics , biochemistry , fluorescence , gene , organic chemistry , physics , polymer , linguistics , philosophy , quantum mechanics
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.