Synthesis of N2-Aryl-2′-Deoxyguanosine Modified Phosphoramidites and Oligonucleotides

Q4 Chemistry
Pratibha P. Ghodke, P. I. Pradeepkumar
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引用次数: 2

Abstract

The N2-position of 2′-deoxyguanosine (N2-position in dG) is well known for forming carcinogenic minor groove DNA adducts, which originate from environmental pollutants, chemicals, and tobacco smoke. The N2-dG DNA adducts have strong implications on biological processes such as DNA replication and repair and may, therefore, result in genomic instability by generating mutations or even cell death. It is crucial to know the role of DNA polymerases when they encounter the N2-dG damaged site in DNA. To get detailed insights on the in vitro DNA damage tolerance or bypass mechanism, there is a need to synthetically access N2-dG damaged DNAs. This article describes a detailed protocol of the synthesis of N2-aryl-dG modified nucleotides using the Buchwald-Hartwig reaction as a main step and incorporation of the modified nucleotides into DNA. In Basic Protocol 1, we focused on the synthesis of five different N2-dG modified phosphoramidites with varying bulkiness (benzyl to pyrenyl). Basic Protocol 2 describes the details of synthesizing N2-dG modified oligonucleotides employing the standard solid phase synthesis protocol. This strategy provides robust synthetic access to various modifications at the N2-position of dG; the modified dGs serve as good substrates to study translesion synthesis and repair pathways. Overall data presented in this article are based on earlier published reports. © 2019 by John Wiley & Sons, Inc.

N2‐芳基‐2′‐脱氧鸟苷修饰的磷酰胺和寡核苷酸的合成
众所周知,2′脱氧鸟苷的N2位(dG中的N2位)会形成致癌的小凹槽DNA加合物,这些加合物来源于环境污染物、化学品和烟草烟雾。N2‐dG DNA加合物对DNA复制和修复等生物学过程具有强烈的影响,因此可能通过产生突变甚至细胞死亡导致基因组不稳定。当DNA聚合酶遇到DNA中N2‐dG受损位点时,了解其作用至关重要。为了深入了解体外DNA损伤耐受性或旁路机制,需要综合获取N2‐dG损伤的DNA。本文描述了以Buchwald-Hartwig反应为主要步骤合成N2‐芳基-dG修饰核苷酸并将修饰核苷酸掺入DNA的详细方案。在基本方案1中,我们重点合成了五种不同的N2‐dG改性的具有不同体积(苄基到芘基)的磷酰胺。基本方案2描述了使用标准固相合成方案合成N2‐dG修饰的寡核苷酸的细节。该策略提供了在dG的N2位置进行各种修饰的稳健合成途径;修饰的dGs是研究跨病变合成和修复途径的良好底物。本文中提供的总体数据基于早期发布的报告。©2019 John Wiley&Sons,股份有限公司版权所有。
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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