Modified Bis-pyrimidine Clamps for Triplex Formation and Their Usein SARS-CoV‑2 Detection

The formation of nucleic acid triple helices (“triplexes”) is an area of great interest due to their potential role in thenatural and artificial regulation of gene expression or for use in analytical, diagnostic, or synthetic methods. During the coronaviruspandemic, a large search for novel methods...

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Autores: Domínguez, Arnau, Gargallo Gómez, Raimundo, Cuestas-Ayllón, Carlos, Gómez-Pinto, Irene, Fàbrega, Carmen , Martínez de la Fuente, Jesús, Damha, M. J., González, Carlos, Eritja i Casadellà, Ramon, Aviñó Andrés, Anna
Formato: artículo
Estado:Versión publicada
Fecha de publicación:2025
País:España
Recursos:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repositorio:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat:2445/227188
Acesso em linha:https://hdl.handle.net/2445/227188
Access Level:acceso abierto
Palavra-chave:Estructura química
Genètica
SARS-CoV-2
Chemical structure
Genetics
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spelling Modified Bis-pyrimidine Clamps for Triplex Formation and Their Usein SARS-CoV‑2 DetectionDomínguez, ArnauGargallo Gómez, RaimundoCuestas-Ayllón, CarlosGómez-Pinto, IreneFàbrega, Carmen Martínez de la Fuente, JesúsDamha, M. J.González, CarlosEritja i Casadellà, RamonAviñó Andrés, AnnaEstructura químicaGenèticaSARS-CoV-2Chemical structureGeneticsSARS-CoV-2The formation of nucleic acid triple helices (“triplexes”) is an area of great interest due to their potential role in thenatural and artificial regulation of gene expression or for use in analytical, diagnostic, or synthetic methods. During the coronaviruspandemic, a large search for novel methods for the detection of SARS-CoV-2 was undertaken. Based on triplex affinity capture andusing polypurine reverse-Hoogsteen hairpins, a method known as Triplex Enhanced Nucleic Acid Detection Assay (TENADA) wasdeveloped for the rapid detection of SARS-CoV-2 without the need for polymerase chain reaction (PCR) amplification. In this work,to expand the targeting scope of this method, we explored triplex-forming bis-pyrimidine clamps targeting a polypurine sequence inthe ORF1a region of SARS-CoV-2. To enhance parallel triplex stability, 2′-sugar and 5-methylpyrimidine modifications wereincorporated into both strands of the clamps, and their effect on the triplexes formed was assessed via NMR and other biophysicalmethods. The results revealed distinct stabilizing effects of the modifications, influenced by their size, sugar puckering, and capacityto form short contacts with neighboring residues. The dual ability of clamps to simultaneously form Watson−Crick and Hoogsteenhydrogen bonds offers a novel perspective on the effect of modifications on triplex stability, previously unexplored with triplexformingoligonucleotides (TFOs). Finally, the bis-pyrimidine clamps that formed the most stable parallel triplexes were applied in athermal lateral flow (TLF) sensing device, demonstrating their potential as biosensing probes. These clamps effectively detected thesynthetic DNA target with limits of detection (LoDs) ranging from 0.05 to 0.001 nM. Understanding the best modification strategiesand their impact on the triplex structure will advance the development of clamps as biosensing and therapeutic agents.American Chemical Society2026202620252026info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersion14 p.application/pdfhttps://hdl.handle.net/2445/227188Articles publicats en revistes (Enginyeria Electrònica i Biomèdica)reponame:Recercat. Dipósit de la Recerca de Catalunyainstname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)InglésReproducció del document publicat a: https://doi.org/10.1021/acsomega.5c02155ACS Omega, 2025, vol. 10, num.22, p. 23535-23548https://doi.org/10.1021/acsomega.5c02155cc-by (c) Domínguez, Arnau et al., 2024https://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:recercat.cat:2445/2271882026-05-29T05:05:01Z
dc.title.none.fl_str_mv Modified Bis-pyrimidine Clamps for Triplex Formation and Their Usein SARS-CoV‑2 Detection
title Modified Bis-pyrimidine Clamps for Triplex Formation and Their Usein SARS-CoV‑2 Detection
spellingShingle Modified Bis-pyrimidine Clamps for Triplex Formation and Their Usein SARS-CoV‑2 Detection
Domínguez, Arnau
Estructura química
Genètica
SARS-CoV-2
Chemical structure
Genetics
SARS-CoV-2
title_short Modified Bis-pyrimidine Clamps for Triplex Formation and Their Usein SARS-CoV‑2 Detection
title_full Modified Bis-pyrimidine Clamps for Triplex Formation and Their Usein SARS-CoV‑2 Detection
title_fullStr Modified Bis-pyrimidine Clamps for Triplex Formation and Their Usein SARS-CoV‑2 Detection
title_full_unstemmed Modified Bis-pyrimidine Clamps for Triplex Formation and Their Usein SARS-CoV‑2 Detection
title_sort Modified Bis-pyrimidine Clamps for Triplex Formation and Their Usein SARS-CoV‑2 Detection
dc.creator.none.fl_str_mv Domínguez, Arnau
Gargallo Gómez, Raimundo
Cuestas-Ayllón, Carlos
Gómez-Pinto, Irene
Fàbrega, Carmen 
Martínez de la Fuente, Jesús
Damha, M. J.
González, Carlos
Eritja i Casadellà, Ramon
Aviñó Andrés, Anna
author Domínguez, Arnau
author_facet Domínguez, Arnau
Gargallo Gómez, Raimundo
Cuestas-Ayllón, Carlos
Gómez-Pinto, Irene
Fàbrega, Carmen 
Martínez de la Fuente, Jesús
Damha, M. J.
González, Carlos
Eritja i Casadellà, Ramon
Aviñó Andrés, Anna
author_role author
author2 Gargallo Gómez, Raimundo
Cuestas-Ayllón, Carlos
Gómez-Pinto, Irene
Fàbrega, Carmen 
Martínez de la Fuente, Jesús
Damha, M. J.
González, Carlos
Eritja i Casadellà, Ramon
Aviñó Andrés, Anna
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Estructura química
Genètica
SARS-CoV-2
Chemical structure
Genetics
SARS-CoV-2
topic Estructura química
Genètica
SARS-CoV-2
Chemical structure
Genetics
SARS-CoV-2
description The formation of nucleic acid triple helices (“triplexes”) is an area of great interest due to their potential role in thenatural and artificial regulation of gene expression or for use in analytical, diagnostic, or synthetic methods. During the coronaviruspandemic, a large search for novel methods for the detection of SARS-CoV-2 was undertaken. Based on triplex affinity capture andusing polypurine reverse-Hoogsteen hairpins, a method known as Triplex Enhanced Nucleic Acid Detection Assay (TENADA) wasdeveloped for the rapid detection of SARS-CoV-2 without the need for polymerase chain reaction (PCR) amplification. In this work,to expand the targeting scope of this method, we explored triplex-forming bis-pyrimidine clamps targeting a polypurine sequence inthe ORF1a region of SARS-CoV-2. To enhance parallel triplex stability, 2′-sugar and 5-methylpyrimidine modifications wereincorporated into both strands of the clamps, and their effect on the triplexes formed was assessed via NMR and other biophysicalmethods. The results revealed distinct stabilizing effects of the modifications, influenced by their size, sugar puckering, and capacityto form short contacts with neighboring residues. The dual ability of clamps to simultaneously form Watson−Crick and Hoogsteenhydrogen bonds offers a novel perspective on the effect of modifications on triplex stability, previously unexplored with triplexformingoligonucleotides (TFOs). Finally, the bis-pyrimidine clamps that formed the most stable parallel triplexes were applied in athermal lateral flow (TLF) sensing device, demonstrating their potential as biosensing probes. These clamps effectively detected thesynthetic DNA target with limits of detection (LoDs) ranging from 0.05 to 0.001 nM. Understanding the best modification strategiesand their impact on the triplex structure will advance the development of clamps as biosensing and therapeutic agents.
publishDate 2025
dc.date.none.fl_str_mv 2025
2026
2026
2026
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/2445/227188
url https://hdl.handle.net/2445/227188
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Reproducció del document publicat a: https://doi.org/10.1021/acsomega.5c02155
ACS Omega, 2025, vol. 10, num.22, p. 23535-23548
https://doi.org/10.1021/acsomega.5c02155
dc.rights.none.fl_str_mv cc-by (c) Domínguez, Arnau et al., 2024
https://creativecommons.org/licenses/by/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv cc-by (c) Domínguez, Arnau et al., 2024
https://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 14 p.
application/pdf
dc.publisher.none.fl_str_mv American Chemical Society
publisher.none.fl_str_mv American Chemical Society
dc.source.none.fl_str_mv Articles publicats en revistes (Enginyeria Electrònica i Biomèdica)
reponame:Recercat. Dipósit de la Recerca de Catalunya
instname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
instname_str Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
reponame_str Recercat. Dipósit de la Recerca de Catalunya
collection Recercat. Dipósit de la Recerca de Catalunya
repository.name.fl_str_mv
repository.mail.fl_str_mv
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