TY  - JOUR
AU  - Kolb, Thorsten
AU  - Khalid, Umar
AU  - Simović, Milena
AU  - Ratnaparkhe, Manasi
AU  - Wong, John
AU  - Jauch, Anna
AU  - Schmezer, Peter
AU  - Rode, Agata
AU  - Sebban, Shulamit
AU  - Haag, Daniel
AU  - Hergt, Michaela
AU  - Devens, Frauke
AU  - Buganim, Yosef
AU  - Zapatka, Marc
AU  - Lichter, Peter
AU  - Ernst, Aurélie
TI  - A versatile system to introduce clusters of genomic double-strand breaks in large cell populations.
JO  - Genes, chromosomes & cancer
VL  - 60
IS  - 5
SN  - 1098-2264
CY  - New York, NY
PB  - Wiley-Liss
M1  - DKFZ-2020-01562
SP  - 303-313
PY  - 2021
N1  - #EA:B420#LA:B420#2021 May;60(5):303-313
AB  - In vitro assays for clustered DNA lesions will facilitate the analysis of the mechanisms underlying complex genome rearrangements such as chromothripsis, including the recruitment of repair factors to sites of DNA double-strand breaks. We present a novel method generating localized DNA double-strand breaks using UV-irradiation with photomasks. The size of the damage foci and the spacing between lesions are fully adjustable, making the assay suitable for different cell types and targeted areas. We validated this set-up with genomically stable epithelial cells, normal fibroblasts, pluripotent stem cells and patient-derived primary cultures. Our method does not require a specialized device such as a laser, making it accessible to a broad range of users. Sensitization by BrdU incorporation is not required, which enables analyzing the DNA damage response in post-mitotic cells. Irradiated cells can be cultivated further, followed by time-lapse imaging or used for downstream biochemical analyses, thanks to the high-throughput of the system. Importantly, we showed genome rearrangements in the irradiated cells, providing a proof of principle for the induction of structural variants by localized DNA lesions. This article is protected by copyright. All rights reserved.
LB  - PUB:(DE-HGF)16
C6  - pmid:32734664
DO  - DOI:10.1002/gcc.22890
UR  - https://inrepo02.dkfz.de/record/157333
ER  -