Interlinked DNA nano-circles for measuring topoisomerase II activity at the level of single decatenation events

TY - JOUR

T1 - Interlinked DNA nano-circles for measuring topoisomerase II activity at the level of single decatenation events

AU - Kristoffersen, Emil L

AU - Jørgensen, Asger Givskov

AU - Jørgensen, Line A

AU - Jensen, Pia W

AU - W Byl, Jo Ann

AU - Osheroff, Neil

AU - Andersen, Anni H

AU - Stougaard, Magnus

AU - Ho, Yi-Ping

AU - Knudsen, Birgitta R

N1 - © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

PY - 2017/7/27

Y1 - 2017/7/27

N2 - DNA nano-structures present appealing new means for monitoring different molecules. Here, we demonstrate the assembly and utilization of a surfaceattached double-stranded DNA catenane composed of two intact interlinked DNA nano-circles for specific and sensitive measurements of the life essential topoisomerase II (Topo II) enzyme activity. Topo II activity was detected via the numeric release of DNA nano-circles, which were visualized at the singlemolecule level in a fluorescence microscope upon isothermal amplification and fluorescence labeling. The transition of each enzymatic reaction to a micrometer sized labeled product enabled quantitative detection of Topo II activity at the single decatenation event level rendering activity measurements in extracts from as few as five cells possible. Topo II activity is a suggested predictive marker in cancer therapy and, consequently, the described highly sensitive monitoring of Topo II activitymay add considerably to the toolbox of individualized medicine where decisions are based on very sparse samples.

AB - DNA nano-structures present appealing new means for monitoring different molecules. Here, we demonstrate the assembly and utilization of a surfaceattached double-stranded DNA catenane composed of two intact interlinked DNA nano-circles for specific and sensitive measurements of the life essential topoisomerase II (Topo II) enzyme activity. Topo II activity was detected via the numeric release of DNA nano-circles, which were visualized at the singlemolecule level in a fluorescence microscope upon isothermal amplification and fluorescence labeling. The transition of each enzymatic reaction to a micrometer sized labeled product enabled quantitative detection of Topo II activity at the single decatenation event level rendering activity measurements in extracts from as few as five cells possible. Topo II activity is a suggested predictive marker in cancer therapy and, consequently, the described highly sensitive monitoring of Topo II activitymay add considerably to the toolbox of individualized medicine where decisions are based on very sparse samples.

KW - Journal Article

UR - http://www.scopus.com/inward/record.url?scp=85026471703&partnerID=8YFLogxK

U2 - 10.1093/nar/gkx480

DO - 10.1093/nar/gkx480

M3 - Journal article

C2 - 28541438

SN - 0305-1048

VL - 45

SP - 7855

EP - 7869

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 13

ER -