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一种经过改进的基于4'-氨甲基三氧杂蒽酮的核酸交联剂,用于双链DNA或RNA的生物素化。

An improved 4'-aminomethyltrioxsalen-based nucleic acid crosslinker for biotinylation of double-stranded DNA or RNA.

作者信息

Wielenberg Kevin, Wang Miao, Yang Min, Ozer Abdullah, Lis John T, Lin Hening

机构信息

Department of Chemistry and Chemical Biology, Cornell University Ithaca NY 14853 USA

Department of Molecular Biology and Genetics, Cornell University Ithaca NY 14853 USA.

出版信息

RSC Adv. 2020 Nov 2;10(65):39870-39874. doi: 10.1039/d0ra07437c. eCollection 2020 Oct 27.

Abstract

Nucleic acid crosslinkers that covalently join complementary strands of DNA/RNA have applications in both pharmaceuticals and as biochemical probes. Psoralen is a popular crosslinking moiety that reacts with double stranded DNA and RNA upon exposure to longwave UV light. The commercially available compound EZ-link psoralen-PEG3-biotin has been used in numerous studies to crosslink DNA and double-stranded RNA for genome-wide investigations. Here we present a new probe, AP3B, which uses the psoralen derivative, 4'-aminomethyltrioxsalen, to crosslink and biotinylate nucleic acids. We show that AP3B is 4 to 5 times more effective at labeling DNA in cells and produces a comparable number of crosslinks with over 100 times less compound and less exposure to UV light than EZ-link psoralen-PEG3-biotin.

摘要

能将DNA/RNA的互补链共价连接起来的核酸交联剂在制药和作为生化探针方面都有应用。补骨脂素是一种常用的交联部分,在暴露于长波紫外光时会与双链DNA和RNA发生反应。市售化合物EZ-link补骨脂素-聚乙二醇3-生物素已在众多研究中用于交联DNA和双链RNA以进行全基因组研究。在此,我们展示了一种新的探针AP3B,它使用补骨脂素衍生物4'-氨甲基三氧杂蒽来交联核酸并进行生物素化。我们表明,AP3B在标记细胞中的DNA方面比EZ-link补骨脂素-聚乙二醇3-生物素有效4至5倍,并且在化合物用量减少100多倍且紫外光暴露更少的情况下能产生相当数量的交联。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/da05/9057442/2aad07217c2c/d0ra07437c-f1.jpg

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