Many methods of nucleic acid labelling are available, which are generally based on linking a specific tag to a sugar phosphate backbone at either 3′- or 5′-end of a nucleic acid molecule. The conjugation of nucleic acids to a ligand, whether it is a biomolecule or a solid support, is brought by the reaction of particular functional groups between the two targets. Thus, a precise knowledge of the contacts made by the transiently binding proteins and nucleic acid molecules is essential to unravel the mechanisms of protein-nucleic acid interaction as well as to understand the architecture of such assemblies. In addition, nucleic acid bioconjugation plays a significant role in elucidating many of cellular processes, mainly those involving protein-DNA/RNA interactions 13. Due to such a broad application range of DNA-based systems, a site-specific immobilization of DNA molecules and subsequent hybridization are the two major events to be developed and optimized. As such, oligonucleotide (ON) bioconjugates hold promise for targeted drug delivery 3, 4, bioimaging 5, 6, biosensing 7, 8, as well as for the development of DNA nanomaterials 9, 10 and DNA-based nanodevices 11, 12. Our results establish novel N 4-cytosine nucleobase modifications as photoreactive labels and suggest an effortless approach for photoimmobilization of nucleic acids.īioconjugation of nucleic acids with diverse biomolecules as well as a variety of solid ligands is of great importance in medicine and bionanotechnology 1, 2. Furthermore, we demonstrate that such immobilized DNA probes can be further used for successful hybridization of complementary DNA targets. Our findings reveal that 3′-tailed single-stranded DNA bearing AP/BP-moieties is easily photoimmobilized onto untreated polystyrene, polypropylene, polylactate, polydimethylsiloxane, sol-gel and borosilicate glass substrates. ![]() Moreover, we show that AP/BP-functionalization of nucleic acid molecules induces an efficient cross-linking upon exposure to UVA light. We find that terminal deoxynucleotidyl transferase-mediated 3′-tailing using AP/BP-containing modified nucleotides generates photoactive DNA, suitable for a straightforward covalent cross-linking with both interacting proteins and a variety of well-known solid polymeric supports. Here, we demonstrate the utilization of N 4-cytidine modified oligonucleotides, which contain reactive acetophenone (AP) or benzophenone (BP) groups, for the UV-induced cross-linking. Since base-modified nucleic acids contribute not only to a broad range of biotechnological fields but also to the understanding of various cellular processes, it is crucial to design novel modifications with unique properties. Bioconjugation, biosensing, bioimaging, bionanomaterials, etc., are only a few examples of application of functionalized DNA.
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