Light‐Activated Control of Translation by Enzymatic Covalent mRNA Labeling
Abstract
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‐queuosine1 (preQ1) 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.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Feb 14, 2018
- Source ID
- 10.1002/anie.201710917
Entities
People
- Cun Yu Zhou
- Dongyan Zhang
- Kayla N. Busby
- Neal Devaraj
- Seth C. Alexander
Organizations
- National Institutes of Health
- United States Department of Defense
- University of California, San Diego