Simple and fast DNA based sensor system for screening of small-molecule compounds targeting eukaryotic topoisomerase 1

Kamilla Vandsø Petersen; Asier Selas; Kirstine Mejlstrup Hymøller; Karol Mizielinski; Maria Thorsager; Magnus Stougaard; Concepcion Alonso; Francisco Palacios; Yolanda Pérez-Pertejo; Rosa M. Reguera; Rafael Balaña-Fouce; Birgitta R. Knudsen; Cinzia Tesauro

doi:10.3390/pharmaceutics13081255

Simple and fast DNA based sensor system for screening of small-molecule compounds targeting eukaryotic topoisomerase 1

Kamilla Vandsø Petersen, Asier Selas, Kirstine Mejlstrup Hymøller, Karol Mizielinski, Maria Thorsager, Magnus Stougaard, Concepcion Alonso, Francisco Palacios, Yolanda Pérez-Pertejo, Rosa M. Reguera, Rafael Balaña-Fouce, Birgitta R. Knudsen, Cinzia Tesauro^*

^*Corresponding author af dette arbejde

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

4 Citationer (Scopus)

Abstract

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 topoisomer-ase 1 targeting drugs, therefore, should involve the possibility of dissecting which step of topoiso-merase 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-mol-ecule 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. Con-clusion: REEAD (C|L) allows for easy and quantitative investigations of topoisomerase 1 targeting compounds and can be performed in non-specialized laboratories.

Originalsprog	Engelsk
Artikelnummer	1255
Tidsskrift	Pharmaceutics
Vol/bind	13
Nummer	8
Antal sider	15
ISSN	1999-4923
DOI	https://doi.org/10.3390/pharmaceutics13081255
Status	Udgivet - aug. 2021

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10.3390/pharmaceutics13081255Licens: CC BY

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Petersen, K. V., Selas, A., Hymøller, K. M., Mizielinski, K., Thorsager, M., Stougaard, M., Alonso, C., Palacios, F., Pérez-Pertejo, Y., Reguera, R. M., Balaña-Fouce, R., Knudsen, B. R., & Tesauro, C. (2021). Simple and fast DNA based sensor system for screening of small-molecule compounds targeting eukaryotic topoisomerase 1. Pharmaceutics, 13(8), Artikel 1255. https://doi.org/10.3390/pharmaceutics13081255

@article{df7fe4e3ec704c0480caf3bbb37be17f,

title = "Simple and fast DNA based sensor system for screening of small-molecule compounds targeting eukaryotic topoisomerase 1",

abstract = "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 topoisomer-ase 1 targeting drugs, therefore, should involve the possibility of dissecting which step of topoiso-merase 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-mol-ecule 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. Con-clusion: REEAD (C|L) allows for easy and quantitative investigations of topoisomerase 1 targeting compounds and can be performed in non-specialized laboratories.",

keywords = "Colorimetric readout, Drug screening, Enzyme activity, REEAD, Rolling circle amplification, Small-molecule compounds, Topoisomerase 1",

author = "Petersen, {Kamilla Vands{\o}} and Asier Selas and Hym{\o}ller, {Kirstine Mejlstrup} and Karol Mizielinski and Maria Thorsager and Magnus Stougaard and Concepcion Alonso and Francisco Palacios and Yolanda P{\'e}rez-Pertejo and Reguera, {Rosa M.} and Rafael Bala{\~n}a-Fouce and Knudsen, {Birgitta R.} and Cinzia Tesauro",

year = "2021",

month = aug,

doi = "10.3390/pharmaceutics13081255",

language = "English",

volume = "13",

journal = "Pharmaceutics",

issn = "1999-4923",

publisher = "Multidisciplinary Digital Publishing Institute (MDPI)",

number = "8",

}

Petersen, KV, Selas, A, Hymøller, KM, Mizielinski, K, Thorsager, M, Stougaard, M, Alonso, C, Palacios, F, Pérez-Pertejo, Y, Reguera, RM, Balaña-Fouce, R, Knudsen, BR & Tesauro, C 2021, 'Simple and fast DNA based sensor system for screening of small-molecule compounds targeting eukaryotic topoisomerase 1', Pharmaceutics, bind 13, nr. 8, 1255. https://doi.org/10.3390/pharmaceutics13081255

Simple and fast DNA based sensor system for screening of small-molecule compounds targeting eukaryotic topoisomerase 1. / Petersen, Kamilla Vandsø; Selas, Asier; Hymøller, Kirstine Mejlstrup et al.
I: Pharmaceutics, Bind 13, Nr. 8, 1255, 08.2021.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

TY - JOUR

T1 - Simple and fast DNA based sensor system for screening of small-molecule compounds targeting eukaryotic topoisomerase 1

AU - Petersen, Kamilla Vandsø

AU - Selas, Asier

AU - Hymøller, Kirstine Mejlstrup

AU - Mizielinski, Karol

AU - Thorsager, Maria

AU - Stougaard, Magnus

AU - Alonso, Concepcion

AU - Palacios, Francisco

AU - Pérez-Pertejo, Yolanda

AU - Reguera, Rosa M.

AU - Balaña-Fouce, Rafael

AU - Knudsen, Birgitta R.

AU - Tesauro, Cinzia

PY - 2021/8

Y1 - 2021/8

N2 - 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 topoisomer-ase 1 targeting drugs, therefore, should involve the possibility of dissecting which step of topoiso-merase 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-mol-ecule 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. Con-clusion: REEAD (C|L) allows for easy and quantitative investigations of topoisomerase 1 targeting compounds and can be performed in non-specialized laboratories.

AB - 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 topoisomer-ase 1 targeting drugs, therefore, should involve the possibility of dissecting which step of topoiso-merase 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-mol-ecule 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. Con-clusion: REEAD (C|L) allows for easy and quantitative investigations of topoisomerase 1 targeting compounds and can be performed in non-specialized laboratories.

KW - Colorimetric readout

KW - Drug screening

KW - Enzyme activity

KW - REEAD

KW - Rolling circle amplification

KW - Small-molecule compounds

KW - Topoisomerase 1

UR - http://www.scopus.com/inward/record.url?scp=85113320400&partnerID=8YFLogxK

U2 - 10.3390/pharmaceutics13081255

DO - 10.3390/pharmaceutics13081255

M3 - Journal article

C2 - 34452216

AN - SCOPUS:85113320400

SN - 1999-4923

VL - 13

JO - Pharmaceutics

JF - Pharmaceutics

IS - 8

M1 - 1255

ER -

Simple and fast DNA based sensor system for screening of small-molecule compounds targeting eukaryotic topoisomerase 1

Abstract

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