Title |
A versatile method for the UVA-induced cross-linking of acetophenone-or benzophenone-functionalized DNA / |
Authors |
Jakubovska, Jevgenija ; Tauraitė, Daiva ; Meškys, Rolandas |
DOI |
10.1038/s41598-018-34892-9 |
Full Text |
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Is Part of |
Scientific reports.. London : Nature Publishing Group. 2018, vol. 8, art. no. 16484, p. [1-10].. ISSN 2045-2322 |
Keywords [eng] |
terminal deoxynucleotidyl transferase ; protein-interactions ; amino-acid ; linked nucleotides ; deoxyribonucleotidyl transferase ; polymerase-lambda ; mass-spectrometry ; drug-delivery ; living cells ; nanomaterials |
Abstract [eng] |
Bioconjugation, biosensing, bioimaging, bionanomaterials, etc., are only a few examples of application of functionalized DNA. 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. 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. 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. Moreover, we show that AP/BP-functionalization of nucleic acid molecules induces an efficient cross-linking upon exposure to UVA light. 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. Furthermore, we demonstrate that such immobilized DNA probes can be further used for successful hybridization of complementary DNA targets. Our results establish novel N-4-cytosine nucleobase modifications as photoreactive labels and suggest an effortless approach for photoimmobilization of nucleic acids. |
Published |
London : Nature Publishing Group |
Type |
Journal article |
Language |
English |
Publication date |
2018 |