3′‐O‐Caged 2′‐Deoxynucleoside Triphosphates for Light‐Mediated, Enzyme‐Catalyzed, Template‐Independent DNA Synthesis. Issue 1 (13th February 2018)

• Record Type: Journal Article
• Authors: Mathews, Anu Stella; Yang, Haikang; Montemagno, Carlo
• Editors: Beaucage, Serge L.; Bergstrom, Donald E.; Herdewijn, Piet; Matsuda, Akira
• Is Part Of: Current protocols in nucleic acid chemistry. Volume 71:Issue 1(2017)
• Page Start: 13.17.1
• Page End: 13.17.38
• Publication Date: 2018-02-13
• Subjects: photo‐cleavable nucleotides -- template independent DNA synthesis -- terminal deoxynucleotidyl transferase -- TdT enzyme; Nucleic acids -- Laboratory manuals; Nucleic Acids -- chemical synthesis; Nucleic acids; Laboratory Manuals; Electronic books; Laboratory manuals; 572.8028
• DOI: 10.1002/cpnc.41
• Languages: English
• ISSNs: 1934-9270

Abstract

Abstract: Synthesis, purification, and characterization of 3′‐ O ‐caged 2′‐deoxyribonucleoside triphosphates (dNTPs), namely 3′‐ O ‐(2‐nitrobenzyl)‐2′‐deoxy ribonucleoside triphosphates (NB‐dNTPs) and 3′‐ O ‐(4, 5‐dimethoxy‐2‐nitrobenzyl)‐2′‐deoxy ribonucleoside triphosphates (DMNB‐dNTPs), are discussed in detail. A total of eight 3′‐ O ‐caged dNTPs are synthesized with specific protocols depending on the nitrogenous base on the first carbon, i.e., adenine, guanine, thymine, and cytosine, as well as the photo‐cleavable group, i.e, 2‐nitrobenzyl and 4, 5‐ dimethoxy‐2‐nitrobenzyl, to be attached in the 3′‐ O position. The purification of the synthesized compounds is done using ion‐exchange and flash chromatography; this is followed by structural confirmation by nuclear magnetic resonance (NMR) and mass spectroscopy (MS). The efficiency of the designed compounds is tested by conducting and evaluating UV‐cleaving experiments at 365 nm with proton NMR and LC‐MS curves. Finally, the application of the 3′‐ O ‐cagged dNTPs in template‐independent, enzyme‐catalyzed, photo‐mediated oligonucleotide synthesis is demonstrated. © 2017 by John Wiley & Sons, Inc.