3,5-二氟-4-甲氧基苄基咪唑啉酮衍生物修饰的2 ' -脱氧尿嘧啶的合成及其用于HER2乳腺癌标志物检测的寡核苷酸探针

Q4 Chemistry
Abed Saady, Noam Y. Steinman, Melissa Wojtyniak, Christian Ducho, Bilha Fischer
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引用次数: 2

摘要

核苷插层偶联物(NICs)描述了我们研究小组开发的一种创新方法,用于制备针对特定mRNA序列(例如乳腺癌标记物)的荧光开启DNA杂交探针。在这种方法中,我们通过柔性间隔将非荧光插入物偶联到核酸(例如尿嘧啶)的碱基上。这种修饰的单体可以通过固相合成并入寡核苷酸中,并且当修饰的寡核苷酸与其互补链杂交时,由于荧光团嵌入在两条链之间,可以观察到较大的荧光增强。5-(6-对甲氧基苄基咪唑啉酮-1-己烯)-2 ' -脱氧尿苷(dUMBI)是将绿色荧光蛋白(GFP)的荧光发色团- 4-甲氧基苄基咪唑啉酮(MBI)通过柔性间隔物偶联而成的一种合成单体。本课题组已经建立了用该探针检测人表皮生长因子受体2 (HER2) mRNA的方法。由于荧光团的自由旋转,单链DNA的荧光强度可以认为是可以忽略不计的。然而,在杂交后,柔性间隔器允许在杂交链之间插入荧光团,从而产生增强的荧光并表明目标mRNA的存在。与MBI荧光团相比,3,5-二氟-4-甲氧基苄基(DFMBI)具有更强的光物理性质。本方案描述了一种简单、可靠、高效、通用的合成改进衍生物dUDFMBI作为荧光开启DNA杂交探针单体的方法,应用于HER2 mRNA的检测。©2020 by John Wiley &基本方案:5-[(6)-3,5-二氟-4-甲氧基苄基咪唑啉-1-己烯]-2 ' -脱氧尿嘧啶的合成
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Synthesis of 2′-Deoxyuridine Modified with a 3,5-Difluoro-4-Methoxybenzylidene Imidazolinone Derivative for Incorporation into Oligonucleotide Probes for Detection of HER2 Breast Cancer Marker

Nucleoside intercalator conjugates (NICs) describe an innovative methodology developed in our research group for preparation of fluorescence turn-on DNA hybridization probes targeting specific mRNA sequences (e.g., breast cancer markers). In this methodology, we conjugate a non-fluorescent intercalator to the base of a nucleic acid (e.g., uracil) via a flexible spacer. This modified monomer can be incorporated into oligonucleotides by solid-phase synthesis and a large fluorescence enhancement is observed when the modified oligonucleotide is hybridized with its complementary strand due to intercalation of the fluorophore between the two strands. 5-(6-p-Methoxybenzylidene imidazolinone-1-hexene)-2′-deoxyuridine (dUMBI) is a synthetic monomer to which 4-methoxybenzylidene imidazolinone (MBI), the fluorescent chromophore of green fluorescent protein (GFP), has been conjugated via a flexible spacer. The detection of human epidermal growth factor receptor 2 (HER2) mRNA by this probe has already been established by our group. The fluorescent intensity of the single-strand DNA can be considered as negligible due to the free rotation of the fluorophore. Upon hybridization, however, the flexible spacer allows for the intercalation of the fluorophore between the hybridized strands, giving rise to enhanced fluorescence and indicating the presence of target mRNA. 3,5-Difluoro-4-methoxybenzylidene (DFMBI) has enhanced photophysical properties compared to MBI fluorophore. This protocol describes a simple, reliable, efficient, and general method for the synthesis of improved derivative dUDFMBI as a monomer of fluorescent turn-on DNA hybridization probe with application for detection of HER2 mRNA. © 2020 by John Wiley & Sons, Inc.

Basic Protocol: Synthesis of 5-[(6)-3,5-difluoro-4-methoxybenzylidene imidazolinone-1-hexene]-2′-deoxyuridine

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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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