[{"data":1,"prerenderedAt":-1},["ShallowReactive",2],{"detail-sidebar-cat-0-en-105":3,"doc-seo-452951-105":59,"doc-detail-452951-en":130},{"code":4,"msg":5,"data":6},0,"success",[7,13,18,23,28,33,38,43,48,51,55],{"id":8,"doc_module":4,"doc_module_name":9,"category_name":10,"show_sort_weight":11,"slug":12},1,"Document","Story & Novel",90,"story-novel",{"id":14,"doc_module":4,"doc_module_name":9,"category_name":15,"show_sort_weight":16,"slug":17},2,"Literature",80,"literature",{"id":19,"doc_module":4,"doc_module_name":9,"category_name":20,"show_sort_weight":21,"slug":22},4,"Exam",70,"exam",{"id":24,"doc_module":4,"doc_module_name":9,"category_name":25,"show_sort_weight":26,"slug":27},5,"Comic",60,"comic",{"id":29,"doc_module":4,"doc_module_name":9,"category_name":30,"show_sort_weight":31,"slug":32},6,"Technology",50,"technology",{"id":34,"doc_module":4,"doc_module_name":9,"category_name":35,"show_sort_weight":36,"slug":37},7,"Healthcare",40,"healthcare",{"id":39,"doc_module":4,"doc_module_name":9,"category_name":40,"show_sort_weight":41,"slug":42},8,"Research & Report",30,"research-report",{"id":44,"doc_module":4,"doc_module_name":9,"category_name":45,"show_sort_weight":46,"slug":47},9,"Religion & Spirituality",20,"religion-spirituality",{"id":46,"doc_module":4,"doc_module_name":9,"category_name":49,"show_sort_weight":46,"slug":50},"World Cup","world-cup",{"id":52,"doc_module":4,"doc_module_name":9,"category_name":53,"show_sort_weight":52,"slug":54},10,"Lifestyle","lifestyle",{"id":56,"doc_module":4,"doc_module_name":9,"category_name":57,"show_sort_weight":24,"slug":58},19,"General","general",{"code":4,"msg":60,"data":61},"ok",{"site_id":62,"language":63,"slug":64,"title":65,"keywords":66,"description":67,"schema_data":68,"social_meta":123,"head_meta":125,"extra_data":127,"updated_unix":129},105,"en","a-modified-x10-23-dnazyme-that-can-better-access-large-structured-rna-targets","A modified X10-23 DNAzyme that can better access large, structured RNA targets","","The 10–23 DNA enzyme is a highly efficient RNA-cleaving DNA catalyst whose recognition arms can be engineered to target AU dinucleotide junctions. Its activity drops on large, structured RNA (lsRNA), such as messenger RNA, even though arm extension, antisense DNA oligonucleotides, and XNA substitutions can partially improve accessibility. Here, multiple XNA patterns with strong RNA binding were introduced into X10-23 recognition arms to enhance lsRNA cleavage. An XdZ-2 variant cleaved selected SARS-CoV-2 lsRNA targets up to 82-fold faster than X10-23, and showed advantages for product release, stability, and low Mg2+ environments.",{"@graph":69,"@context":122},[70,84,105],{"@type":71,"itemListElement":72},"BreadcrumbList",[73,77,79,82],{"item":74,"name":75,"@type":76,"position":8},"https://docshare.wps.com","Home","ListItem",{"item":78,"name":9,"@type":76,"position":14},"https://docshare.wps.com/document/",{"item":80,"name":35,"@type":76,"position":81},"https://docshare.wps.com/document/healthcare/",3,{"item":83,"name":65,"@type":76,"position":19},"https://docshare.wps.com/document/a-modified-x10-23-dnazyme-that-can-better-access-large-structured-rna-targets/452951/",{"url":83,"name":65,"@type":85,"image":86,"author":91,"headline":65,"publisher":94,"fileFormat":97,"inLanguage":63,"description":67,"dateModified":98,"datePublished":99,"encodingFormat":97,"isAccessibleForFree":100,"interactionStatistic":101},"DigitalDocument",{"url":87,"@type":88,"width":89,"height":90},"https://docshare.wps.com/thumbnails/a-modified-x10-23-dnazyme-that-can-better-access-large-structured-rna-targets/452951.png","ImageObject",300,407,{"name":92,"@type":93},"Logic","Person",{"url":74,"name":95,"@type":96},"DocShare","Organization","application/pdf","2026-10-30","2026-09-30",true,{"@type":102,"interactionType":103,"userInteractionCount":19},"InteractionCounter",{"@type":104},"ViewAction",{"@type":106,"mainEntity":107},"FAQPage",[108,114,118],{"name":109,"@type":110,"acceptedAnswer":111},"Why does the 10–23 DNAzyme show reduced activity on large, structured RNA (lsRNA) targets?","Question",{"text":112,"@type":113},"The 10–23 DNAzyme often has reduced accessibility and cleavage efficiency for lsRNA, such as messenger RNA, compared with short unstructured RNA. This limitation is especially evident at lower Mg2+ concentrations.","Answer",{"name":115,"@type":110,"acceptedAnswer":116},"What modification strategy was used to improve lsRNA access in this study?",{"text":117,"@type":113},"XNA patterns with high RNA-binding strength were substituted into the substrate recognition arms of X10-23. The work focuses on designing specific XNA arm sequences to enhance target accessibility.",{"name":119,"@type":110,"acceptedAnswer":120},"What performance did the XdZ-2 enzyme achieve compared with X10-23?",{"text":121,"@type":113},"XdZ-2, featuring a distinct 2′-F-RNA-LNA-FANA arm pattern, could access several lsRNA targets from SARS-CoV-2 and achieved cleavage rates up to 82-fold faster than X10-23 in one system.","https://schema.org",{"og:url":83,"og:type":124,"og:title":65,"og:site_name":95,"og:description":67},"article",{"robots":126,"canonical":83},"index,follow",{"doc_id":128,"site_id":62},452951,1790795898,{"code":4,"msg":5,"data":131},{"doc_id":128,"user_id":132,"nickname":92,"user_avatar":133,"doc_module":4,"category_id":34,"category_name":35,"doc_title":65,"doc_description":67,"doc_content":134,"file_id":135,"file_url":136,"file_type":137,"file_size":138,"view_count":19,"is_deleted":4,"is_public":8,"is_downloadable":8,"audit_status":8,"page_count":139,"language":140,"language_code":63,"site_id":62,"html_lang":63,"table_of_contents":141,"faqs":142,"seo_title":143,"seo_description":67,"update_tm":144,"read_time":145,"preview_page_count":4},1099513958762,"https://ap-avatar.wpscdn.com/avatar/1000023916a998db790?x-image-process=image/resize,m_fixed,w_180,h_180&k=1784791008015729253","A modified X10-23 DNAzyme that can better access large, structured RNA targets  \nConnor Nurmi 1,2 , Halle M. Barber3 , Harneesh Kaur3 , John D. Brennan 2 , Masad J. Damha 3 ,* , Yingfu Li 1 ,*  \n1 Department of Biochemistry and Biomedical Sciences, McMaster University, Ontario L8S 4L8, Canada  \n2 Biointerfaces Institute, McMaster University, Ontario L8S 4L8, Canada  \n3 Department of Chemistry, McGill University, Montreal, Quebec H3A 0G4, Canada  \n∗ To whom correspondence should be addressed. Email: [liying@mcmaster.ca](liying@mcmaster.ca)  \nCorrespondence may also be addressed to Masad [J. Damha. Email:](J. Damha. Email: masad.damha@mcgill.ca)[ masad.damha@mcgill.ca](J. Damha. Email: masad.damha@mcgill.ca)  \nAbstract  \nThe 10–23 DNA enzyme is one of the most efficient RNA-cleaving enzymes reported, possessing substrate recognition arms that can be designed to target virtually any AU diribonucleotide junction. However, 10–23 often shows reduced activity for large, structured RNA (lsRNA) substrates like messenger RNA. Increasing arm length or adding antisense DNA oligonucleotides (ASOs) can improve accessibility to lsRNA but may also reduce the efficiency of product release. Xeno nucleic acids (XNAs), such as 2􀀃-fluoro-arabinonucleic acid (FANA), have been substituted for DNA into the arms of 10–23 to improve activity, such as in the FANA-modified X10-23, but X10-23 also shows poor accessibility for lsRNA targets. To overcome this issue, we substituted patterns of various XNAs with high RNA binding strength into the substrate recognition arms of X10-23 . We found that an X10-23 enzyme with a distinct 2􀀃 F-RNA-LNA-FANA arm pattern, denoted as XdZ-2, could gain access to several lsRNA targets from SARS-CoV-2, achieving cleavage rates up to 82-fold faster than X10-23 for one system. While the ASO strategy provided higher cleavage rates for two other lsRNA systems, XdZ-2 may be a more attractive alternative in low Mg2+ environments and in terms of improving the efficiency of product release and stability in biological samples.  \nGraphical abstract  \nIntroduction  \nThe ‘10–23’DNA enzyme, also known as 10–23 DNAzyme, 10–23, or simply dZ, is a single-stranded DNA oligonucleotide that catalyzes the sequence-specific trans cleavage of RNA, employing modifiable substrate recognition arms that flank a central, 15-nt-long, catalytic core sequence [1] . 10– 23 was developed by in vitro selection and remains one of the most efficient RNA-cleaving DNAzymes reported [2, 3]. How-  \never, the rate of RNA transesterification by 10–23 still falls behind protein enzymes and RNA-based catalysts (ribozymes), and as a result, many groups have explored various strategies to improve its activity.  \nSubstitution of xeno nucleic acids (XNAs) into the substrate recognition arms and/or catalytic core of the 10–23 DNAzyme has been explored by other groups to improve RNA-cleavage activity [4, 5] . These studies primarily used  \nReceived: June 15, 2025. Revised: November 10, 2025. Accepted: November 25, 2025 © The Author(s) 2026. Published by Oxford University Press.  \nThis is an Open Access article distributed under the terms of the Creative Commons Attribution License ([https://creativecommons.org/licenses/by/4.0/](https://creativecommons.org/licenses/by/4.0/)), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.  \n2 Nurmi et al.  \nXNAs with modifications to the 2􀀃 position of the ribose sugar, demonstrating improved RNA heteroduplex stability compared to RNA:RNA and DNA:RNA duplexes. The first XNA to be substituted into the substrate recognition arms of 10–23 was 2􀀃 -OMe, which aimed to prevent exonucleolytic degradation in biological samples [6] . Since then, 2􀀃 -OMe [7, 8] and other XNAs, such as locked nucleic acids (LNA)[9–12] and 2􀀃 -fluoro-arabinonucleic acids (FANA) [4], have been explored for their ability to improve the RNA-cleavage activity of 10–23. The development of polyme","cbCairyZcpZGgZgS","https://ap.wps.com/l/cbCairyZcpZGgZgS","pdf",3112463,13,"English","# Abstract\n# Introduction\n## Background on the 10–23 DNA enzyme\n## Prior XNA and arm-substitution strategies\n## Limitations with lsRNA and Mg2+ dependence","[{\"question\":\"Why does the 10–23 DNAzyme show reduced activity on large, structured RNA (lsRNA) targets?\",\"answer\":\"The 10–23 DNAzyme often has reduced accessibility and cleavage efficiency for lsRNA, such as messenger RNA, compared with short unstructured RNA. This limitation is especially evident at lower Mg2+ concentrations.\"},{\"question\":\"What modification strategy was used to improve lsRNA access in this study?\",\"answer\":\"XNA patterns with high RNA-binding strength were substituted into the substrate recognition arms of X10-23. The work focuses on designing specific XNA arm sequences to enhance target accessibility.\"},{\"question\":\"What performance did the XdZ-2 enzyme achieve compared with X10-23?\",\"answer\":\"XdZ-2, featuring a distinct 2′-F-RNA-LNA-FANA arm pattern, could access several lsRNA targets from SARS-CoV-2 and achieved cleavage rates up to 82-fold faster than X10-23 in one system.\"}]","A modified X10-23 DNAzyme that can better access large, structured RNA targets | PDF",1790737933,33]