Real-time detection of TDP1 activity using a fluorophore-quencher coupled DNA-biosensor

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Standard

Real-time detection of TDP1 activity using a fluorophore-quencher coupled DNA-biosensor. / Jensen, Pia Wrensted; Falconi, Mattia; Kristoffersen, Emil Laust; Simonsen, Anita Tranberg; Cifuentes, Jéssica B.; Marcussen, Lærke; Hougaard, Rikke Frøhlich; Vagner, Josephine; Harmsen, Charlotte; Jensen, Sissel Juul; Ho, Yi-Ping; Withers, Marjorie A.; Lupski, James R.; Koch, Jørn Erland; Desideri, Alessandro; Knudsen, Birgitta R.; Stougaard, Magnus.

I: Biosensors and Bioelectronics, Bind Volume 48, 15.10.2013, s. 230–237.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

Harvard

Jensen, PW, Falconi, M, Kristoffersen, EL, Simonsen, AT, Cifuentes, JB, Marcussen, L, Hougaard, RF, Vagner, J, Harmsen, C, Jensen, SJ, Ho, Y-P, Withers, MA, Lupski, JR, Koch, JE, Desideri, A, Knudsen, BR & Stougaard, M 2013, 'Real-time detection of TDP1 activity using a fluorophore-quencher coupled DNA-biosensor', Biosensors and Bioelectronics, bind Volume 48, s. 230–237. https://doi.org/10.1016/j.bios.2013.04.019

APA

Jensen, P. W., Falconi, M., Kristoffersen, E. L., Simonsen, A. T., Cifuentes, J. B., Marcussen, L., Hougaard, R. F., Vagner, J., Harmsen, C., Jensen, S. J., Ho, Y-P., Withers, M. A., Lupski, J. R., Koch, J. E., Desideri, A., Knudsen, B. R., & Stougaard, M. (2013). Real-time detection of TDP1 activity using a fluorophore-quencher coupled DNA-biosensor. Biosensors and Bioelectronics, Volume 48, 230–237. https://doi.org/10.1016/j.bios.2013.04.019

CBE

Jensen PW, Falconi M, Kristoffersen EL, Simonsen AT, Cifuentes JB, Marcussen L, Hougaard RF, Vagner J, Harmsen C, Jensen SJ, Ho Y-P, Withers MA, Lupski JR, Koch JE, Desideri A, Knudsen BR, Stougaard M. 2013. Real-time detection of TDP1 activity using a fluorophore-quencher coupled DNA-biosensor. Biosensors and Bioelectronics. Volume 48:230–237. https://doi.org/10.1016/j.bios.2013.04.019

MLA

Jensen, Pia Wrensted o.a.. "Real-time detection of TDP1 activity using a fluorophore-quencher coupled DNA-biosensor". Biosensors and Bioelectronics. 2013, Volume 48. 230–237. https://doi.org/10.1016/j.bios.2013.04.019

Vancouver

Jensen PW, Falconi M, Kristoffersen EL, Simonsen AT, Cifuentes JB, Marcussen L o.a. Real-time detection of TDP1 activity using a fluorophore-quencher coupled DNA-biosensor. Biosensors and Bioelectronics. 2013 okt 15;Volume 48:230–237. https://doi.org/10.1016/j.bios.2013.04.019

Author

Jensen, Pia Wrensted ; Falconi, Mattia ; Kristoffersen, Emil Laust ; Simonsen, Anita Tranberg ; Cifuentes, Jéssica B. ; Marcussen, Lærke ; Hougaard, Rikke Frøhlich ; Vagner, Josephine ; Harmsen, Charlotte ; Jensen, Sissel Juul ; Ho, Yi-Ping ; Withers, Marjorie A. ; Lupski, James R. ; Koch, Jørn Erland ; Desideri, Alessandro ; Knudsen, Birgitta R. ; Stougaard, Magnus. / Real-time detection of TDP1 activity using a fluorophore-quencher coupled DNA-biosensor. I: Biosensors and Bioelectronics. 2013 ; Bind Volume 48. s. 230–237.

Bibtex

@article{54ce99b2c1b448829c45bd5512b56ab2,
title = "Real-time detection of TDP1 activity using a fluorophore-quencher coupled DNA-biosensor",
abstract = "Real-time detection of enzyme activities may present the easiest and most reliable way of obtaining quantitative analyses in biological samples. We present a new DNA-biosensor capable of detecting the activity of the potential anticancer drug target tyrosyl-DNA phosphodiesterase 1 (TDP1) in a very simple, high throughput, and real-time format. The biosensor is specific for Tdp1 even in complex biological samples, such as human cell extracts, and may consequently find future use in fundamental studies as well as a cancer predictive tool allowing fast analyses of diagnostic cell samples such as biopsies. TDP1 removes covalent 3′DNA adducts in DNA single-strand break repair. This enzymatic activity forms the basis of the design of the TDP1-biosensor, which consists of a short hairpin-forming oligonucleotide having a 5′fluorophore and a 3′quencher brought in close proximity by the secondary structure of the biosensor. The specific action of TDP1 removes the quencher, thereby enabling optical detection of the fluorophore. Since the enzymatic action of TDP1 is the only “signal amplification” the increase in fluorescence may easily be followed in real-time and allows quantitative analyses of TDP1 activity in pure enzyme fractions as well as in crude cell extracts. In the present study we demonstrate the specificity of the biosensor, its ability to quantitatively detect up- or down-regulated TDP1 activity, and that it may be used for measuring and for analyzing the mechanism of TDP1 inhibition.",
keywords = " Tyrosyl-DNA phosphodiesterase 1 (TDP1), Biosensor, Enzyme activity, Real-time measurement, Fluorophore–quencher",
author = "Jensen, {Pia Wrensted} and Mattia Falconi and Kristoffersen, {Emil Laust} and Simonsen, {Anita Tranberg} and Cifuentes, {J{\'e}ssica B.} and L{\ae}rke Marcussen and Hougaard, {Rikke Fr{\o}hlich} and Josephine Vagner and Charlotte Harmsen and Jensen, {Sissel Juul} and Yi-Ping Ho and Withers, {Marjorie A.} and Lupski, {James R.} and Koch, {J{\o}rn Erland} and Alessandro Desideri and Knudsen, {Birgitta R.} and Magnus Stougaard",
year = "2013",
month = oct,
day = "15",
doi = "10.1016/j.bios.2013.04.019",
language = "English",
volume = "Volume 48",
pages = "230–237",
journal = "Biosensors and Bioelectronics",
issn = "0956-5663",
publisher = "Elsevier BV",

}

RIS

TY - JOUR

T1 - Real-time detection of TDP1 activity using a fluorophore-quencher coupled DNA-biosensor

AU - Jensen, Pia Wrensted

AU - Falconi, Mattia

AU - Kristoffersen, Emil Laust

AU - Simonsen, Anita Tranberg

AU - Cifuentes, Jéssica B.

AU - Marcussen, Lærke

AU - Hougaard, Rikke Frøhlich

AU - Vagner, Josephine

AU - Harmsen, Charlotte

AU - Jensen, Sissel Juul

AU - Ho, Yi-Ping

AU - Withers, Marjorie A.

AU - Lupski, James R.

AU - Koch, Jørn Erland

AU - Desideri, Alessandro

AU - Knudsen, Birgitta R.

AU - Stougaard, Magnus

PY - 2013/10/15

Y1 - 2013/10/15

N2 - Real-time detection of enzyme activities may present the easiest and most reliable way of obtaining quantitative analyses in biological samples. We present a new DNA-biosensor capable of detecting the activity of the potential anticancer drug target tyrosyl-DNA phosphodiesterase 1 (TDP1) in a very simple, high throughput, and real-time format. The biosensor is specific for Tdp1 even in complex biological samples, such as human cell extracts, and may consequently find future use in fundamental studies as well as a cancer predictive tool allowing fast analyses of diagnostic cell samples such as biopsies. TDP1 removes covalent 3′DNA adducts in DNA single-strand break repair. This enzymatic activity forms the basis of the design of the TDP1-biosensor, which consists of a short hairpin-forming oligonucleotide having a 5′fluorophore and a 3′quencher brought in close proximity by the secondary structure of the biosensor. The specific action of TDP1 removes the quencher, thereby enabling optical detection of the fluorophore. Since the enzymatic action of TDP1 is the only “signal amplification” the increase in fluorescence may easily be followed in real-time and allows quantitative analyses of TDP1 activity in pure enzyme fractions as well as in crude cell extracts. In the present study we demonstrate the specificity of the biosensor, its ability to quantitatively detect up- or down-regulated TDP1 activity, and that it may be used for measuring and for analyzing the mechanism of TDP1 inhibition.

AB - Real-time detection of enzyme activities may present the easiest and most reliable way of obtaining quantitative analyses in biological samples. We present a new DNA-biosensor capable of detecting the activity of the potential anticancer drug target tyrosyl-DNA phosphodiesterase 1 (TDP1) in a very simple, high throughput, and real-time format. The biosensor is specific for Tdp1 even in complex biological samples, such as human cell extracts, and may consequently find future use in fundamental studies as well as a cancer predictive tool allowing fast analyses of diagnostic cell samples such as biopsies. TDP1 removes covalent 3′DNA adducts in DNA single-strand break repair. This enzymatic activity forms the basis of the design of the TDP1-biosensor, which consists of a short hairpin-forming oligonucleotide having a 5′fluorophore and a 3′quencher brought in close proximity by the secondary structure of the biosensor. The specific action of TDP1 removes the quencher, thereby enabling optical detection of the fluorophore. Since the enzymatic action of TDP1 is the only “signal amplification” the increase in fluorescence may easily be followed in real-time and allows quantitative analyses of TDP1 activity in pure enzyme fractions as well as in crude cell extracts. In the present study we demonstrate the specificity of the biosensor, its ability to quantitatively detect up- or down-regulated TDP1 activity, and that it may be used for measuring and for analyzing the mechanism of TDP1 inhibition.

KW - Tyrosyl-DNA phosphodiesterase 1 (TDP1)

KW - Biosensor

KW - Enzyme activity

KW - Real-time measurement

KW - Fluorophore–quencher

U2 - 10.1016/j.bios.2013.04.019

DO - 10.1016/j.bios.2013.04.019

M3 - Journal article

C2 - 23693093

VL - Volume 48

SP - 230

EP - 237

JO - Biosensors and Bioelectronics

JF - Biosensors and Bioelectronics

SN - 0956-5663

ER -