Loading…

Orthogonal End Labelling of Oligonucleotides through Dual Incorporation of Click‐Reactive NTP Analogues

Post‐synthetic modification of nucleic acid structures with clickable functionality is a versatile tool that facilitates many emerging applications, including immune evasion, enhancements in stability, fluorescent labelling, chemical 5′‐RNA‐capping and the development of functional aptamers. While c...

Full description

Saved in:
Bibliographic Details
Published in:Chembiochem : a European journal of chemical biology 2024-01, Vol.25 (1), p.e202300701-n/a
Main Authors: Schönegger, Eva S., Crisp, Antony, Radukic, Marco, Burmester, Jonas, Frischmuth, Thomas, Carell, Thomas
Format: Article
Language:English
Subjects:
Citations: Items that this one cites
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Post‐synthetic modification of nucleic acid structures with clickable functionality is a versatile tool that facilitates many emerging applications, including immune evasion, enhancements in stability, fluorescent labelling, chemical 5′‐RNA‐capping and the development of functional aptamers. While certain chemoenzymatic approaches for 3′‐azido and alkynyl labelling are known, equivalent 5′‐strategies are either inefficient, complex, or require harsh chemical conditions. Here, we present a modular and facile technology to consecutively modify DNA and RNA strands at both ends with click‐modifiable functional groups. Our approach using γ‐modified ATP analogues facilitates T4 PNK‐catalysed 5′‐modification of oligonucleotides, a process that is compatible with TdT‐catalysed 3′‐elongation using 3′‐azido‐2′,3′‐ddGTP. Finally, we demonstrate that our approach is suitable for both oligo‐oligo ligations, as well ssDNA circularization. We anticipate that such approaches will pave the way for the synthesis of highly functionalised oligonucleotides, improving the therapeutic and diagnostic applicability of oligonucleotides such as in the realm of next‐generation sequencing. A new chemoenzymatic method involving dual incorporation of γ‐ and 3′‐modified NTPs allows sequential 5′‐ and 3′‐end labelling of DNA and RNA molecules.
ISSN:1439-4227
1439-7633
DOI:10.1002/cbic.202300701