Simple and Fast DNA Based Sensor System for Screening of Small-Molecule Compounds Targeting Eukaryotic Topoisomerase 1

Background: Eukaryotic topoisomerase 1 is a potential target of anti-parasitic and anti-cancer drugs. Parasites require topoisomerase 1 activity for survival and, consequently, compounds that inhibit topoisomerase 1 activity may be of interest. All effective topoisomerase 1 drugs with anti-cancer ac...

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Autores: Petersen, Kamilla Vandsø, Selas Lanseros, Asier, Hymøller, Kirstine Mejlstrup, Mizielinski, Karol, Thorsager, Maria, Stougaard, Magnus, Alonso Pérez, Concepción Estibaliz, Palacios Gambra, Francisco Javier, Pérez-Pertejo, Yolanda, Reguera, Rosa M., Balaña-Fouce, Rafael, Knudsen, Birgitta Ruth, Tesauro, Cinzia
Tipo de recurso: artículo
Fecha de publicación:2021
País:España
Institución:Universidad del País Vasco
Repositorio:Addi. Archivo Digital para la Docencia y la Investigación
OAI Identifier:oai:addi.ehu.eus:10810/53077
Acceso en línea:http://hdl.handle.net/10810/53077
Access Level:acceso abierto
Palabra clave:topoisomerase 1
REEAD
rolling circle amplification
small-molecule compounds
drug screening
enzyme activity
colorimetric readout
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spelling Simple and Fast DNA Based Sensor System for Screening of Small-Molecule Compounds Targeting Eukaryotic Topoisomerase 1Petersen, Kamilla VandsøSelas Lanseros, AsierHymøller, Kirstine MejlstrupMizielinski, KarolThorsager, MariaStougaard, MagnusAlonso Pérez, Concepción EstibalizPalacios Gambra, Francisco JavierPérez-Pertejo, YolandaReguera, Rosa M.Balaña-Fouce, RafaelKnudsen, Birgitta RuthTesauro, Cinziatopoisomerase 1REEADrolling circle amplificationsmall-molecule compoundsdrug screeningenzyme activitycolorimetric readoutBackground: Eukaryotic topoisomerase 1 is a potential target of anti-parasitic and anti-cancer drugs. Parasites require topoisomerase 1 activity for survival and, consequently, compounds that inhibit topoisomerase 1 activity may be of interest. All effective topoisomerase 1 drugs with anti-cancer activity act by inhibiting the ligation reaction of the enzyme. Screening for topoisomerase 1 targeting drugs, therefore, should involve the possibility of dissecting which step of topoisomerase 1 activity is affected. Methods: Here we present a novel DNA-based assay that allows for screening of the effect of small-molecule compounds targeting the binding/cleavage or the ligation steps of topoisomerase 1 catalysis. This novel assay is based on the detection of a rolling circle amplification product generated from a DNA circle resulting from topoisomerase 1 activity. Results: We show that the binding/cleavage and ligation reactions of topoisomerase 1 can be investigated separately in the presented assay termed REEAD (C|L) and demonstrate that the assay can be used to investigate, which of the individual steps of topoisomerase 1 catalysis are affected by small-molecule compounds. The assay is gel-free and the results can be detected by a simple colorimetric readout method using silver-on-gold precipitation rendering large equipment unnecessary. Conclusion: REEAD (C|L) allows for easy and quantitative investigations of topoisomerase 1 targeting compounds and can be performed in non-specialized laboratories.K.V. thanks Aase and Ejnar Danielsens Fond, Aage and Johanne Louis-Hansens Fond and Edel and Wilhelm Daubenmerkls Almenvelgørende Fond; A.S. thanks the Basque Government for a formation contract.MDPI2021202120212021info:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10810/53077reponame:Addi. Archivo Digital para la Docencia y la Investigacióninstname:Universidad del País VascoIngléshttps://www.mdpi.com/1999-4923/13/8/1255/htminfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by/3.0/es/2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).oai:addi.ehu.eus:10810/530772026-06-18T09:23:17Z
dc.title.none.fl_str_mv Simple and Fast DNA Based Sensor System for Screening of Small-Molecule Compounds Targeting Eukaryotic Topoisomerase 1
title Simple and Fast DNA Based Sensor System for Screening of Small-Molecule Compounds Targeting Eukaryotic Topoisomerase 1
spellingShingle Simple and Fast DNA Based Sensor System for Screening of Small-Molecule Compounds Targeting Eukaryotic Topoisomerase 1
Petersen, Kamilla Vandsø
topoisomerase 1
REEAD
rolling circle amplification
small-molecule compounds
drug screening
enzyme activity
colorimetric readout
title_short Simple and Fast DNA Based Sensor System for Screening of Small-Molecule Compounds Targeting Eukaryotic Topoisomerase 1
title_full Simple and Fast DNA Based Sensor System for Screening of Small-Molecule Compounds Targeting Eukaryotic Topoisomerase 1
title_fullStr Simple and Fast DNA Based Sensor System for Screening of Small-Molecule Compounds Targeting Eukaryotic Topoisomerase 1
title_full_unstemmed Simple and Fast DNA Based Sensor System for Screening of Small-Molecule Compounds Targeting Eukaryotic Topoisomerase 1
title_sort Simple and Fast DNA Based Sensor System for Screening of Small-Molecule Compounds Targeting Eukaryotic Topoisomerase 1
dc.creator.none.fl_str_mv Petersen, Kamilla Vandsø
Selas Lanseros, Asier
Hymøller, Kirstine Mejlstrup
Mizielinski, Karol
Thorsager, Maria
Stougaard, Magnus
Alonso Pérez, Concepción Estibaliz
Palacios Gambra, Francisco Javier
Pérez-Pertejo, Yolanda
Reguera, Rosa M.
Balaña-Fouce, Rafael
Knudsen, Birgitta Ruth
Tesauro, Cinzia
author Petersen, Kamilla Vandsø
author_facet Petersen, Kamilla Vandsø
Selas Lanseros, Asier
Hymøller, Kirstine Mejlstrup
Mizielinski, Karol
Thorsager, Maria
Stougaard, Magnus
Alonso Pérez, Concepción Estibaliz
Palacios Gambra, Francisco Javier
Pérez-Pertejo, Yolanda
Reguera, Rosa M.
Balaña-Fouce, Rafael
Knudsen, Birgitta Ruth
Tesauro, Cinzia
author_role author
author2 Selas Lanseros, Asier
Hymøller, Kirstine Mejlstrup
Mizielinski, Karol
Thorsager, Maria
Stougaard, Magnus
Alonso Pérez, Concepción Estibaliz
Palacios Gambra, Francisco Javier
Pérez-Pertejo, Yolanda
Reguera, Rosa M.
Balaña-Fouce, Rafael
Knudsen, Birgitta Ruth
Tesauro, Cinzia
author2_role author
author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv topoisomerase 1
REEAD
rolling circle amplification
small-molecule compounds
drug screening
enzyme activity
colorimetric readout
topic topoisomerase 1
REEAD
rolling circle amplification
small-molecule compounds
drug screening
enzyme activity
colorimetric readout
description Background: Eukaryotic topoisomerase 1 is a potential target of anti-parasitic and anti-cancer drugs. Parasites require topoisomerase 1 activity for survival and, consequently, compounds that inhibit topoisomerase 1 activity may be of interest. All effective topoisomerase 1 drugs with anti-cancer activity act by inhibiting the ligation reaction of the enzyme. Screening for topoisomerase 1 targeting drugs, therefore, should involve the possibility of dissecting which step of topoisomerase 1 activity is affected. Methods: Here we present a novel DNA-based assay that allows for screening of the effect of small-molecule compounds targeting the binding/cleavage or the ligation steps of topoisomerase 1 catalysis. This novel assay is based on the detection of a rolling circle amplification product generated from a DNA circle resulting from topoisomerase 1 activity. Results: We show that the binding/cleavage and ligation reactions of topoisomerase 1 can be investigated separately in the presented assay termed REEAD (C|L) and demonstrate that the assay can be used to investigate, which of the individual steps of topoisomerase 1 catalysis are affected by small-molecule compounds. The assay is gel-free and the results can be detected by a simple colorimetric readout method using silver-on-gold precipitation rendering large equipment unnecessary. Conclusion: REEAD (C|L) allows for easy and quantitative investigations of topoisomerase 1 targeting compounds and can be performed in non-specialized laboratories.
publishDate 2021
dc.date.none.fl_str_mv 2021
2021
2021
2021
dc.type.none.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/10810/53077
url http://hdl.handle.net/10810/53077
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv https://www.mdpi.com/1999-4923/13/8/1255/htm
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by/3.0/es/
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by/3.0/es/
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv reponame:Addi. Archivo Digital para la Docencia y la Investigación
instname:Universidad del País Vasco
instname_str Universidad del País Vasco
reponame_str Addi. Archivo Digital para la Docencia y la Investigación
collection Addi. Archivo Digital para la Docencia y la Investigación
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15.300719

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