Unique Crosslinking Properties of Psoralen‐conjugated Oligonucleotides developed by Novel Psoralen N‐Hydroxysuccinimide Esters
Unique Crosslinking Properties of Psoralen‐conjugated Oligonucleotides developed by Novel Psoralen N‐Hydroxysuccinimide Esters
复制标题
新型补骨脂素 N-羟基琥珀酰亚胺酯开发的补骨脂素缀合寡核苷酸的独特交联特性
DOI：
10.1002/cbic.202200789
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发表时间：
2023
期刊：
影响因子：
3.2
通讯作者：
Yamayoshi Asako
中科院分区：
文献类型：
--
作者：
Nakao Juki;Mikame Yu;Eshima Honoka;Yamamoto Tsuyoshi;Dohno Chikara;Wada Takehiko;Yamayoshi Asako
补骨脂素及其衍生物，如三氧沙林，对DNA具有独特的交联性。然而，补骨脂素单体与靶DNA不具有序列特异性的交联能力。随着补骨脂素偶联寡核苷酸的发展，补骨脂素偶联分子在基因转录抑制、基因敲除和通过基因组编辑进行靶向重组等方面的应用日益广泛。在这项研究中，我们开发了两种新的补骨脂烯N-羟基琥珀酰亚胺(NHS)酯，可以将补骨脂素引入任何氨基修饰的寡核苷酸中。定量评价了Ps-寡糖对单链DNA的光交联效率，结果表明，对5-MC的选择性是Trioxsalen的独特之处。我们发现，在补骨脂素的C-5位引入寡核苷酸接头可以促进目标双链DNA的良好交联。我们相信我们的发现是发展Ps-寡核苷酸作为新的基因调控工具的重要信息。
Psoralens and their derivatives, such as trioxsalen, have unique crosslinking features to DNA. However, psoralen monomers do not have sequence‐specific crosslinking ability with the target DNA. With the development of psoralen‐conjugated oligonucleotides (Ps‐Oligos), sequence‐specific crosslinking with target DNA has become achievable, thereby expanding the application of psoralen‐conjugated molecules in gene transcription inhibition, gene knockout, and targeted recombination by genome editing. In this study, we developed two novel psoralenN‐hydroxysuccinimide (NHS) esters that allow the introduction of psoralens into any amino‐modified oligonucleotides. Quantitative evaluation of the photo‐crosslinking efficiencies of the Ps‐Oligos to target single‐stranded DNAs revealed that the crosslinking selectivity to 5‐mC is the unique feature of trioxsalen. We found that the introduction of an oligonucleotideviaa linker at the C‐5 position of psoralen can promote favorable crosslinking to target double‐stranded DNA. We believe our findings are essential information for the development of Ps‐Oligos as novel gene regulation tools.