Nucleotide-resolution DNA double-strand break mapping by next-generation sequencing.
Nucleotide-resolution DNA double-strand break mapping by next-generation sequencing.
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DOI：
10.1038/nmeth.2408
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发表时间：
2013-04
期刊：
影响因子：
48
通讯作者：
Dikic, Ivan
中科院分区：
文献类型：
--
作者：
Crosetto, Nicola;Mitra, Abhishek;Silva, Maria Joao;Bienko, Magda;Dojer, Norbert;Wang, Qi;Karaca, Elif;Chiarle, Roberto;Skrzypczak, Magdalena;Ginalski, Krzysztof;Pasero, Philippe;Rowicka, Maga;Dikic, Ivan
我们提出了一种全基因组方法，可通过直接原位断裂标记，链霉亲和素的富集和下一代测序（Bless）来绘制核苷酸分辨率DNA双链断裂（DSB）。我们使用不同的人类和小鼠细胞，诱导DSB的剂以及测序平台对Bless进行了全面验证和测试。 Bless能够检测到端粒末端，SCE核酸内切酶引起的DSB和复杂的全基因组DSB景观。作为原则的证明，我们表征了对人类细胞复制应激的敏感性的基因组景观，并鉴定出超过两千种非均匀分布的蚜虫素敏感区域（ASRS）在基因中过度代表性并富含卫星重复。在人类癌症中重新排列的区域也富含ASR，其中许多与癌症相关的基因对复制应激表现出很高的敏感性。我们的方法适用于在各种细胞中DSB的全基因组映射和实验条件，其特异性和分辨率无法通过当前技术实现。
We present a genome-wide method to map DNA double-strand breaks (DSBs) at nucleotide resolution by direct in situ breaks labeling, enrichment on streptavidin, and next-generation sequencing (BLESS). We comprehensively validated and tested BLESS using different human and mouse cells, DSBs-inducing agents, and sequencing platforms. BLESS was able to detect telomere ends, Sce endonuclease-induced DSBs, and complex genome-wide DSBs landscapes. As a proof of principle, we characterized the genomic landscape of sensitivity to replication stress in human cells, and identified over two thousand non-uniformly distributed aphidicolin-sensitive regions (ASRs) overrepresented in genes and enriched in satellite repeats. ASRs were also enriched in regions rearranged in human cancers, with many cancer-associated genes exhibiting high sensitivity to replication stress. Our method is suitable for genome-wide mapping of DSBs in various cells and experimental conditions with a specificity and resolution unachievable by current techniques.
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影响因子：
64.5
作者：
Casper, AM;Nghiem, P;Glover, TW
通讯作者：
Glover, TW
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
21.3
作者：
Feng, Wenyi;Collingwood, David;Boeck, Max E;Fox, Lindsay A;Alvino, Gina M;Fangman, Walton L;Raghuraman, Mosur K;Brewer, Bonita J
通讯作者：
Brewer, Bonita J
DOI：
10.1038/nrc1299
发表时间：
2004-03
期刊：
Nature reviews. Cancer
影响因子：
--
作者：
通讯作者：
--
DOI：
10.1073/pnas.0603779103
发表时间：
2006-06-27
影响因子：
11.1
作者：
Marti, Thomas M.;Hefner, Eli;Cleaver, James E.
通讯作者：
Cleaver, James E.