3′-O-Caged 2′-Deoxynucleoside Triphosphates for Light-Mediated, Enzyme-Catalyzed, Template-Independent DNA Synthesis

Q4 Chemistry
Anu Stella Mathews, Haikang Yang, Carlo Montemagno
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引用次数: 10

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.

3 ' -O-Caged 2 ' -脱氧核苷三磷酸光介导,酶催化,模板非依赖性DNA合成
详细讨论了3 ' - o笼型2 ' -脱氧核糖核苷三磷酸(dNTPs),即3 ' - o -(2-硝基苄基)-2 ' -脱氧核糖核苷三磷酸(NB-dNTPs)和3 ' - o -(4,5-二甲氧基-2-硝基苄基)-2 ' -脱氧核糖核苷三磷酸(DMNB-dNTPs)的合成、纯化和表征。根据第一个碳上的含氮碱基,即腺嘌呤、鸟嘌呤、胸腺嘧啶和胞嘧啶,以及附着在3′- o位置的光可切割基团,即2-硝基苯基和4,5-二甲氧基-2-硝基苯基,以特定的方案合成了总共8个3′- o笼型dNTPs。合成化合物的纯化采用离子交换和闪蒸色谱法;其次是核磁共振(NMR)和质谱(MS)的结构确认。通过质子核磁共振和LC-MS曲线在365 nm处进行紫外切割实验,并对所设计化合物的效率进行了测试。最后,展示了3 ' - o- cage dNTPs在模板不依赖、酶催化、光介导的寡核苷酸合成中的应用。©2017 by John Wiley &儿子,Inc。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Current Protocols in Nucleic Acid Chemistry
Current Protocols in Nucleic Acid Chemistry Chemistry-Organic Chemistry
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期刊介绍: Published in association with International Society for Nucleosides, Nucleotides & Nucleic Acids (IS3NA) , Current Protocols in Nucleic Acid Chemistry is equally valuable for biotech, pharmaceutical, and academic labs. It is the resource for designing and running successful research projects in the rapidly growing and changing field of nucleic acid, nucleotide, and nucleoside research.
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