Specific Monitoring the DNA Helicase Function via Anchor‐Embedded DNA Probe
Abstract DNA helicases play a pivotal role in maintaining genome integrity by unwinding the DNA double helix and are often considered promising targets for drug development. However, assessing specific DNA helicase activity in living cells remains challenging. Herein, the first anchor‐embedded duple...
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| Format: | Article |
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Wiley
2025-02-01
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| Series: | Advanced Science |
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| Online Access: | https://doi.org/10.1002/advs.202413368 |
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| author | Keni Ning Xiaoyan Tang Zhe Li Liting Zhong Yingchen Zhou Jiaen Wang Wanyi Huang Han Zhang Jiajun Ke Tiangang Luan Shuo‐Bin Chen Junqiu Zhai |
| author_facet | Keni Ning Xiaoyan Tang Zhe Li Liting Zhong Yingchen Zhou Jiaen Wang Wanyi Huang Han Zhang Jiajun Ke Tiangang Luan Shuo‐Bin Chen Junqiu Zhai |
| author_sort | Keni Ning |
| collection | DOAJ |
| description | Abstract DNA helicases play a pivotal role in maintaining genome integrity by unwinding the DNA double helix and are often considered promising targets for drug development. However, assessing specific DNA helicase activity in living cells remains challenging. Herein, the first anchor‐embedded duplex (ATED) probe, 17GC, is constructed to uniquely monitor the unwinding activity of Werner syndrome helicase (WRN), a clinical anticancer target. This probe integrates biophysical screening and molecular simulation approaches. The 17GC probe consists of two components: the first one is a bubble structure as an anchor for recruiting WRN in cells, and the second one is GC‐rich double helices on both ends of the bubble, which allow high sensitivity in detecting WRN activity. In vitro evaluations demonstrate that 17GC is highly sensitive and specific to WRN (LOD = 33.5 pm) compared to a wide range of other enzymes, including helicases and nucleases. Cellular evaluation reveals that the ATED probe exhibits remarkable performance in monitoring WRN helicase activity and assessing the inhibitory efficiency of clinical WRN inhibitors in various cell types. This study introduces a novel approach for designing specific and sensitive probes for DNA helicases in cells, which holds promise for biological characterization and drug development. |
| format | Article |
| id | doaj-art-42e147619a31407ca486bbdde6e6bcdd |
| institution | OA Journals |
| issn | 2198-3844 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Wiley |
| record_format | Article |
| series | Advanced Science |
| spelling | doaj-art-42e147619a31407ca486bbdde6e6bcdd2025-08-20T02:30:35ZengWileyAdvanced Science2198-38442025-02-01128n/an/a10.1002/advs.202413368Specific Monitoring the DNA Helicase Function via Anchor‐Embedded DNA ProbeKeni Ning0Xiaoyan Tang1Zhe Li2Liting Zhong3Yingchen Zhou4Jiaen Wang5Wanyi Huang6Han Zhang7Jiajun Ke8Tiangang Luan9Shuo‐Bin Chen10Junqiu Zhai11School of Pharmaceutical Sciences Guangzhou University of Chinese Medicine Guangzhou 510006 ChinaSchool of Pharmaceutical Sciences Guangzhou University of Chinese Medicine Guangzhou 510006 ChinaSchool of Pharmaceutical Sciences, Guangdong Provincial Key Laboratory of New Drug Design and Evaluation Sun Yat‐Sen University Guangzhou 510006 ChinaSchool of Pharmaceutical Sciences, Guangdong Provincial Key Laboratory of New Drug Design and Evaluation Sun Yat‐Sen University Guangzhou 510006 ChinaSchool of Pharmaceutical Sciences, Guangdong Provincial Key Laboratory of New Drug Design and Evaluation Sun Yat‐Sen University Guangzhou 510006 ChinaSchool of Pharmaceutical Sciences, Guangdong Provincial Key Laboratory of New Drug Design and Evaluation Sun Yat‐Sen University Guangzhou 510006 ChinaSchool of Pharmaceutical Sciences, Guangdong Provincial Key Laboratory of New Drug Design and Evaluation Sun Yat‐Sen University Guangzhou 510006 ChinaSchool of Pharmaceutical Sciences Guangzhou University of Chinese Medicine Guangzhou 510006 ChinaSchool of Pharmaceutical Sciences Guangzhou University of Chinese Medicine Guangzhou 510006 ChinaSchool of Environmental and Chemical Engineering Wuyi University Jiangmen 529020 ChinaSchool of Pharmaceutical Sciences, Guangdong Provincial Key Laboratory of New Drug Design and Evaluation Sun Yat‐Sen University Guangzhou 510006 ChinaSchool of Pharmaceutical Sciences Guangzhou University of Chinese Medicine Guangzhou 510006 ChinaAbstract DNA helicases play a pivotal role in maintaining genome integrity by unwinding the DNA double helix and are often considered promising targets for drug development. However, assessing specific DNA helicase activity in living cells remains challenging. Herein, the first anchor‐embedded duplex (ATED) probe, 17GC, is constructed to uniquely monitor the unwinding activity of Werner syndrome helicase (WRN), a clinical anticancer target. This probe integrates biophysical screening and molecular simulation approaches. The 17GC probe consists of two components: the first one is a bubble structure as an anchor for recruiting WRN in cells, and the second one is GC‐rich double helices on both ends of the bubble, which allow high sensitivity in detecting WRN activity. In vitro evaluations demonstrate that 17GC is highly sensitive and specific to WRN (LOD = 33.5 pm) compared to a wide range of other enzymes, including helicases and nucleases. Cellular evaluation reveals that the ATED probe exhibits remarkable performance in monitoring WRN helicase activity and assessing the inhibitory efficiency of clinical WRN inhibitors in various cell types. This study introduces a novel approach for designing specific and sensitive probes for DNA helicases in cells, which holds promise for biological characterization and drug development.https://doi.org/10.1002/advs.202413368DNA helicasesDNA probeintracellular monitoringWerner syndrome helicase |
| spellingShingle | Keni Ning Xiaoyan Tang Zhe Li Liting Zhong Yingchen Zhou Jiaen Wang Wanyi Huang Han Zhang Jiajun Ke Tiangang Luan Shuo‐Bin Chen Junqiu Zhai Specific Monitoring the DNA Helicase Function via Anchor‐Embedded DNA Probe Advanced Science DNA helicases DNA probe intracellular monitoring Werner syndrome helicase |
| title | Specific Monitoring the DNA Helicase Function via Anchor‐Embedded DNA Probe |
| title_full | Specific Monitoring the DNA Helicase Function via Anchor‐Embedded DNA Probe |
| title_fullStr | Specific Monitoring the DNA Helicase Function via Anchor‐Embedded DNA Probe |
| title_full_unstemmed | Specific Monitoring the DNA Helicase Function via Anchor‐Embedded DNA Probe |
| title_short | Specific Monitoring the DNA Helicase Function via Anchor‐Embedded DNA Probe |
| title_sort | specific monitoring the dna helicase function via anchor embedded dna probe |
| topic | DNA helicases DNA probe intracellular monitoring Werner syndrome helicase |
| url | https://doi.org/10.1002/advs.202413368 |
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