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Light‐Activated Control of Translation by Enzymatic Covalent mRNA Labeling
Author(s) -
Zhang Dongyang,
Zhou Cun Yu,
Busby Kayla N.,
Alexander Seth C.,
Devaraj Neal K.
Publication year - 2018
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201710917
Subject(s) - messenger rna , translation (biology) , untranslated region , guanine , rna , gene expression , chemistry , transfer rna , biochemistry , protein biosynthesis , riboswitch , nucleotide , gene , microbiology and biotechnology , biology , non coding rna
Activation of cellular protein expression upon visible‐light photocleavage of small‐molecule caging groups covalently attached to the 5′ untranslated region (5′ UTR) of an mRNA was achieved. These photocleavable caging groups are conjugated to in vitro transcribed mRNA (IVT‐mRNA) through RNA transglycosylation, an enzymatic process in which a bacterial tRNA guanine transglycosylase (TGT) exchanges a guanine nucleobase in a specific 17‐nucleotide motif (Tag) for synthetic pre‐queuosine 1 (preQ 1 ) derivatives. The caging groups severely reduce mRNA translation efficiency when strategically placed in the 5′ UTR. Using this method, we demonstrate the successful spatiotemporal photoregulation of gene expression with single‐cell precision. Our method can be applied to therapeutically relevant chemically modified mRNA (mod‐mRNA) transcripts. This strategy provides a modular and efficient approach for developing synthetic gene regulatory circuits, biotechnological applications, and therapeutic discovery.