Journal Article

DKFZ-2020-01562

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png A versatile system to introduce clusters of genomic double-strand breaks in large cell populations.

Kolb, T. (First author)DKFZ* ; Khalid, U.DKFZ* ; Simović, M.DKFZ* ; Ratnaparkhe, M.DKFZ* ; Wong, J.DKFZ* ; Jauch, A. ; Schmezer, P.DKFZ* ; Rode, A.DKFZ* ; Sebban, S. ; Haag, D.DKFZ* ; Hergt, M.DKFZ* ; Devens, F.DKFZ* ; Buganim, Y. ; Zapatka, M.DKFZ* ; Lichter, P.DKFZ* ; Ernst, A. (Last author)DKFZ*

2021
Wiley-Liss New York, NY

This record in other databases:  

Please use a persistent id in citations: doi:10.1002/gcc.22890

Abstract: 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.

Note: #EA:B420#LA:B420#2021 May;60(5):303-313

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Contributing Institute(s):

1. Genominstabilität in Tumoren (B420)
2. DKTK HD zentral (HD01)
3. Epigenomik (B370)
4. B062 Pädiatrische Neuroonkologie (B062)
5. B060 Molekulare Genetik (B060)

Research Program(s):

1. 312 - Funktionelle und strukturelle Genomforschung (POF4-312) (POF4-312)

Appears in the scientific report 2021

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Database coverage:
; BIOSIS Previews ; Biological Abstracts ; Clarivate Analytics Master Journal List ; Current Contents - Life Sciences ; DEAL Wiley ; Essential Science Indicators ; IF < 5 ; JCR ; NCBI Molecular Biology Database ; Nationallizenz ; SCOPUS ; Science Citation Index ; Science Citation Index Expanded ; Web of Science Core Collection

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