Design and synthesis of fluorescence-labeled nucleotide with a cleavable aryl-triazene linker for DNA sequencing by synthesis

Nucleosides Nucleotides Nucleic Acids. 2024 Nov 18:1-16. doi: 10.1080/15257770.2024.2429696. Online ahead of print.

Abstract

A novel acid-cleavable triazene linker was synthesized and then reacted with modified 2'-deoxyuridine triphosphate (dUTP), followed by Cy3 NHS ester, the final product serves as an excellent reversible terminator for DNA sequencing by synthesis (DNA SBS). The synthesized dye-labeled terminator incorporated into DNA strand faithfully in a DNA-elongation, and the fluorophore incorporated into DNA strands was cleaved completely under weak acidic conditions within short time. Further the first cleaved product can be incorporated with 100% efficiency for the second time. These preliminary evaluations show that the triazene reversible terminator has a great potential value in DNA sequencing.

Keywords: DNA sequencing by synthesis; Triazene linker; reversible terminator; triphosphate analogues.