Yifan Gao, Wanqin Zhang, Jingyi Si, Xizhong Shen, Qunyan Yao and Changfeng Zhu
{"title":"T7 RNA聚合酶†产生的孔雀石绿适体对坝甲基转移酶活性的无标记检测及抑制作用","authors":"Yifan Gao, Wanqin Zhang, Jingyi Si, Xizhong Shen, Qunyan Yao and Changfeng Zhu","doi":"10.1039/D4NJ05217J","DOIUrl":null,"url":null,"abstract":"<p >DNA methyltransferase (DNA MTase) catalyzes DNA methylation, which plays a crucial role in various biological processes such as chromatin assembly, aging, and tumorigenesis. For this reason, developing handy DNA MTase assays and screening its inhibitors have potential value in clinical applications. Here, we have established a label-free strategy for Dam MTase, using the T7 RNA transcription system for signal amplification and its transcription product malachite green aptamer (MGA) for signal transduction. This strategy reached a detection limit of 9.95 × 10<small><sup>−5</sup></small> U mL<small><sup>−1</sup></small> and successfully achieved detection in a complex biological matrix. Simultaneously, this method has realized the screening of methyltransferase inhibitors, laying a solid foundation for its clinical application.</p>","PeriodicalId":95,"journal":{"name":"New Journal of Chemistry","volume":" 10","pages":" 4218-4225"},"PeriodicalIF":2.5000,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Label-free detection of dam methyltransferase activity and inhibition via malachite green aptamer generated by T7 RNA polymerase†\",\"authors\":\"Yifan Gao, Wanqin Zhang, Jingyi Si, Xizhong Shen, Qunyan Yao and Changfeng Zhu\",\"doi\":\"10.1039/D4NJ05217J\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p >DNA methyltransferase (DNA MTase) catalyzes DNA methylation, which plays a crucial role in various biological processes such as chromatin assembly, aging, and tumorigenesis. For this reason, developing handy DNA MTase assays and screening its inhibitors have potential value in clinical applications. Here, we have established a label-free strategy for Dam MTase, using the T7 RNA transcription system for signal amplification and its transcription product malachite green aptamer (MGA) for signal transduction. This strategy reached a detection limit of 9.95 × 10<small><sup>−5</sup></small> U mL<small><sup>−1</sup></small> and successfully achieved detection in a complex biological matrix. Simultaneously, this method has realized the screening of methyltransferase inhibitors, laying a solid foundation for its clinical application.</p>\",\"PeriodicalId\":95,\"journal\":{\"name\":\"New Journal of Chemistry\",\"volume\":\" 10\",\"pages\":\" 4218-4225\"},\"PeriodicalIF\":2.5000,\"publicationDate\":\"2025-02-19\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"New Journal of Chemistry\",\"FirstCategoryId\":\"92\",\"ListUrlMain\":\"https://pubs.rsc.org/en/content/articlelanding/2025/nj/d4nj05217j\",\"RegionNum\":3,\"RegionCategory\":\"化学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"CHEMISTRY, MULTIDISCIPLINARY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"New Journal of Chemistry","FirstCategoryId":"92","ListUrlMain":"https://pubs.rsc.org/en/content/articlelanding/2025/nj/d4nj05217j","RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"CHEMISTRY, MULTIDISCIPLINARY","Score":null,"Total":0}
引用次数: 0
摘要
DNA甲基转移酶(DNA MTase)催化DNA甲基化,在染色质组装、衰老和肿瘤发生等多种生物过程中起着至关重要的作用。因此,开发方便的DNA MTase检测方法并筛选其抑制剂具有潜在的临床应用价值。在这里,我们建立了一种无标记的Dam MTase策略,使用T7 RNA转录系统进行信号扩增,其转录产物孔雀石绿适体(malachite green aptamer, MGA)进行信号转导。该策略达到了9.95 × 10−5 U mL−1的检出限,并成功地在复杂的生物基质中实现了检测。同时该方法实现了甲基转移酶抑制剂的筛选,为其临床应用奠定了坚实的基础。
Label-free detection of dam methyltransferase activity and inhibition via malachite green aptamer generated by T7 RNA polymerase†
DNA methyltransferase (DNA MTase) catalyzes DNA methylation, which plays a crucial role in various biological processes such as chromatin assembly, aging, and tumorigenesis. For this reason, developing handy DNA MTase assays and screening its inhibitors have potential value in clinical applications. Here, we have established a label-free strategy for Dam MTase, using the T7 RNA transcription system for signal amplification and its transcription product malachite green aptamer (MGA) for signal transduction. This strategy reached a detection limit of 9.95 × 10−5 U mL−1 and successfully achieved detection in a complex biological matrix. Simultaneously, this method has realized the screening of methyltransferase inhibitors, laying a solid foundation for its clinical application.