Comment on: “Atom‐Modified gDNA Enhances Cleavage Activity of TtAgo Enabling Ultrasensitive Nucleic Acid Testing”
Abstract The recent article by Zhang et al. piqued the interest. An atom‐modification‐based strategy is reported to enhance the cleavage activity of TtAgo, improving its practicability in TtAgo‐based nucleic acid testing. Specifically, the 2′‐fluorine (2′F)‐modified guide DNA (2′F‐gDNA) shows signif...
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| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2025-04-01
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| Series: | Advanced Science |
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| Online Access: | https://doi.org/10.1002/advs.202406872 |
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| author | Yue Tang Xiao‐han Wang Xu Xu Lu‐sheng Xin Xiu‐dan Wang Xin‐min Li |
| author_facet | Yue Tang Xiao‐han Wang Xu Xu Lu‐sheng Xin Xiu‐dan Wang Xin‐min Li |
| author_sort | Yue Tang |
| collection | DOAJ |
| description | Abstract The recent article by Zhang et al. piqued the interest. An atom‐modification‐based strategy is reported to enhance the cleavage activity of TtAgo, improving its practicability in TtAgo‐based nucleic acid testing. Specifically, the 2′‐fluorine (2′F)‐modified guide DNA (2′F‐gDNA) shows significant enhancement in the cleavage activity of TtAgo on double‐stranded (dsDNA) by increasing the melting temperature (Tm) and strengthening the binding affinity between 2′F‐gDNA and the targeted dsDNA. These findings are considered important for both molecular diagnostics and gene editing. A careful review of the article, however, raises questions that merit further discussion. After comprehensively reviewing the cleavage mechanism and structure of TtAgo‐gDNA‐target ternary complexes, and thoroughly analyzing our results, it is believed that the increased Tm and binding affinity of 2′F‐gDNA are not the primary factors that enhance cleavage activity, it is speculated that the 2′F modification at gDNA 3′‐end likely influences the propagation step. The data suggest that several details need to be addressed to improve the robustness of 2′F‐gDNA/TtAgo cleavage. |
| format | Article |
| id | doaj-art-e03f5306b9124bd58ba3dd112c59d54a |
| institution | DOAJ |
| issn | 2198-3844 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Wiley |
| record_format | Article |
| series | Advanced Science |
| spelling | doaj-art-e03f5306b9124bd58ba3dd112c59d54a2025-08-20T03:16:56ZengWileyAdvanced Science2198-38442025-04-011213n/an/a10.1002/advs.202406872Comment on: “Atom‐Modified gDNA Enhances Cleavage Activity of TtAgo Enabling Ultrasensitive Nucleic Acid Testing”Yue Tang0Xiao‐han Wang1Xu Xu2Lu‐sheng Xin3Xiu‐dan Wang4Xin‐min Li5Institute of chronic diseases，Institute for Translational Medicine The Affiliated Hospital of Qingdao University School of Basic Medicine，College of Medicine Qingdao University Qingdao 266073 ChinaInstitute of chronic diseases，Institute for Translational Medicine The Affiliated Hospital of Qingdao University School of Basic Medicine，College of Medicine Qingdao University Qingdao 266073 ChinaState Key Laboratory of Marine Food Processing & Safety Control College of Food Science and Engineering Ocean University of China Qingdao 266404 ChinaJining Medical University Jining 272067 ChinaState Key Laboratory of Marine Food Processing & Safety Control College of Food Science and Engineering Ocean University of China Qingdao 266404 ChinaInstitute of chronic diseases，Institute for Translational Medicine The Affiliated Hospital of Qingdao University School of Basic Medicine，College of Medicine Qingdao University Qingdao 266073 ChinaAbstract The recent article by Zhang et al. piqued the interest. An atom‐modification‐based strategy is reported to enhance the cleavage activity of TtAgo, improving its practicability in TtAgo‐based nucleic acid testing. Specifically, the 2′‐fluorine (2′F)‐modified guide DNA (2′F‐gDNA) shows significant enhancement in the cleavage activity of TtAgo on double‐stranded (dsDNA) by increasing the melting temperature (Tm) and strengthening the binding affinity between 2′F‐gDNA and the targeted dsDNA. These findings are considered important for both molecular diagnostics and gene editing. A careful review of the article, however, raises questions that merit further discussion. After comprehensively reviewing the cleavage mechanism and structure of TtAgo‐gDNA‐target ternary complexes, and thoroughly analyzing our results, it is believed that the increased Tm and binding affinity of 2′F‐gDNA are not the primary factors that enhance cleavage activity, it is speculated that the 2′F modification at gDNA 3′‐end likely influences the propagation step. The data suggest that several details need to be addressed to improve the robustness of 2′F‐gDNA/TtAgo cleavage.https://doi.org/10.1002/advs.2024068722'‐fluorine modified gDNAdsDNA cleavagegene editingnucleic acid detectionTtAgo |
| spellingShingle | Yue Tang Xiao‐han Wang Xu Xu Lu‐sheng Xin Xiu‐dan Wang Xin‐min Li Comment on: “Atom‐Modified gDNA Enhances Cleavage Activity of TtAgo Enabling Ultrasensitive Nucleic Acid Testing” Advanced Science 2'‐fluorine modified gDNA dsDNA cleavage gene editing nucleic acid detection TtAgo |
| title | Comment on: “Atom‐Modified gDNA Enhances Cleavage Activity of TtAgo Enabling Ultrasensitive Nucleic Acid Testing” |
| title_full | Comment on: “Atom‐Modified gDNA Enhances Cleavage Activity of TtAgo Enabling Ultrasensitive Nucleic Acid Testing” |
| title_fullStr | Comment on: “Atom‐Modified gDNA Enhances Cleavage Activity of TtAgo Enabling Ultrasensitive Nucleic Acid Testing” |
| title_full_unstemmed | Comment on: “Atom‐Modified gDNA Enhances Cleavage Activity of TtAgo Enabling Ultrasensitive Nucleic Acid Testing” |
| title_short | Comment on: “Atom‐Modified gDNA Enhances Cleavage Activity of TtAgo Enabling Ultrasensitive Nucleic Acid Testing” |
| title_sort | comment on atom modified gdna enhances cleavage activity of ttago enabling ultrasensitive nucleic acid testing |
| topic | 2'‐fluorine modified gDNA dsDNA cleavage gene editing nucleic acid detection TtAgo |
| url | https://doi.org/10.1002/advs.202406872 |
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