Quantification of single-strand DNA lesions caused by the topoisomerase II poison etoposide using single DNA molecule imaging
Artikel i vetenskaplig tidskrift, 2022

DNA-damaging agents, such as radiation and chemotherapy, are common in cancer treatment, but the dosing has proven to be challenging, leading to severe side effects in some patients. Hence, to be able to personalize DNA-damaging chemotherapy, it is important to develop fast and reliable methods to measure the resulting DNA damage in patient cells. Here, we demonstrate how single DNA molecule imaging using fluorescence microscopy can quantify DNA-damage caused by the topoisomerase II (TopoII) poison etoposide. The assay uses an enzyme cocktail consisting of base excision repair (BER) enzymes to repair the DNA damage caused by etoposide and label the sites using a DNA polymerase and fluorescently labeled nucleotides. Using this DNA-damage detection assay we find a large variation in etoposide induced DNA-damage after in vitro treatment of blood cells from healthy individuals. We furthermore used the TopoII inhibitor ICRF-193 to show that the etoposide-induced damage in DNA was TopoII dependent. We discuss how our results support a potential future use of the assay for personalized dosing of chemotherapy.

Base excision repair

DNA damage

Nick translation

Single molecule imaging


Single-strand breaks


Vandana Singh

Chalmers, Biologi och bioteknik, Kemisk biologi

Sahlgrenska universitetssjukhuset

Pegah Johansson

Sahlgrenska universitetssjukhuset

Göteborgs universitet

Elina Ekedahl

Yii Lih Lin

Chalmers, Biologi och bioteknik, Kemisk biologi

Ola Hammarsten

Göteborgs universitet

Sahlgrenska universitetssjukhuset

Fredrik Westerlund

Chalmers, Biologi och bioteknik, Kemisk biologi

Biochemical and Biophysical Research Communications

0006-291X (ISSN) 1090-2104 (eISSN)

Vol. 594 57-62


Nanovetenskap och nanoteknik

Hälsa och teknik


Annan medicinsk grundvetenskap

Farmakologi och toxikologi

Medicinsk bioteknologi (med inriktning mot cellbiologi (inklusive stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci)





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