Synthesis of 5'-NAD-Capped RNA

Höfer, K.; Abele, F.; Schlotthauer, J.; Jäschke, A.

doi:10.1021/acs.bioconjchem.6b00072

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Synthesis of 5'-NAD-Capped RNA

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https://www.ncbi.nlm.nih.gov/pubmed/26942556
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Höfer, K., Abele, F., Schlotthauer, J., & Jäschke, A. (2016). Synthesis of 5'-NAD-Capped RNA. Bioconjug Chem, 27(4), 874-7. doi:10.1021/acs.bioconjchem.6b00072.

Cite as: https://hdl.handle.net/21.11116/0000-000A-6B0F-F

Abstract

In prokaryotic organisms, certain regulatory RNAs have recently been found to be linked to the ubiquitous redox cofactor nicotinamide adenine dinucleotide (NAD) at their 5'-ends. Biochemical and structural investigations of this new caplike RNA modification require synthetic access to pure NAD-RNA. Here we report a chemoenzymatic approach to generate 5'-NAD-capped RNA in high yields and purity under mild conditions. This approach uses unprotected 5'-monophosphate RNA synthesized either chemically or enzymatically, 5',5'-pyrophosphate bond formation by phosphorimidazolide chemistry, and an enzymatic cleanup step. Thus, 5'-NAD-modified RNA can be synthesized independent of length, structure, and nucleotide sequence.