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Bioorthogonal site-specific labeling of the 5'-cap structure in eukaryotic mRNAs.
A chemo-enzymatic approach for site-specific labeling of 5'-capped RNAs based on bioorthogonal chemistry is presented and further modified using strain-promoted azide-alkyne cycloaddition (SPAAC). Expand
A chemo-enzymatic approach for site-specific modification of the RNA cap.
A two-step approach can be used to site-specifically modify the 5'-cap of eukaryotic mRNAs to introduce bioorthogonal groups using S-adenosyl-L-methionine-based cosubstrates. Expand
Enzymatic modification of 5′-capped RNA with a 4-vinylbenzyl group provides a platform for photoclick and inverse electron-demand Diels–Alder reaction† †Electronic supplementary information (ESI)
Enzymatic transfer of 4-vinylbenzyl to the mRNA 5′-cap gives access to the fluorogenic photoclick and the inverse electron-demand Diels–Alder reaction.
Modifying the 5'-Cap for Click Reactions of Eukaryotic mRNA and To Tune Translation Efficiency in Living Cells.
The efficient and versatile N7 manipulation of the mRNA cap makes mRNAs amenable to both modulation of their biological function and intracellular labeling, and represents a valuable addition to the chemical biology toolbox. Expand
Current covalent modification methods for detecting RNA in fixed and living cells.
This review focuses on covalent labeling approaches that rely on the introduction of a small reactive group into the nascent or completed transcript followed by bioorthogonal click chemistry. Expand
Engineering Giardia lamblia trimethylguanosine synthase (GlaTgs2) to transfer non-natural modifications to the RNA 5'-cap.
This work reports on engineering GlaTgs2 to enhance the activity on AdoPropen, and introduces the best substitution at the homologous position led to variant T34A with ∼40-fold higher specificity for AdOPropen than the original Gla Tgs2 WT. Expand
Dual 5' Cap Labeling Based on Regioselective RNA Methyltransferases and Bioorthogonal Reactions.
This work has presented a straightforward enzymatic approach to allow site-specific modification of two different positions on the 5' cap of eukaryotic mRNA with either identical or different small functional groups, and it can be extended to a three-enzyme cascade reaction for their in situ production. Expand
cation of 5 0-capped RNA with a 4-vinylbenzyl group provides a platform for photoclick and inverse electron-demand Diels – Alder reaction †
Chemo-enzymatic strategies provide a highly selective means to label different classes of biomolecules in vitro, but also in vivo. In the field of RNA, efficient labeling of eukaryotic mRNA withExpand
High-Throughput, Lysis-free Screening for Sulfatase Activity Using Escherichia coli Autodisplay in Microdroplets.
In SpAS1 variants with improved performance toward the bulky fluorescein disulfate, many of the beneficial mutations occur in residues that form hydrogen bonds between α-helices in the C-terminal oligomerization region, suggesting a previously unknown role for the dimer interface in shaping the substrate binding site of SpAS2. Expand
Enzymatic Modification of 5'-Capped RNA and Subsequent Labeling by Click Chemistry.
This work describes a chemoenzymatic method to site specifically label 5'-capped model mRNAs independent of their sequence, engineered to introduce alkyne, azido, or 4-vinylbenzyl moieties to the 5'-cap. Expand