Biernacka Anna, Zhu Yingjie, Skrzypczak Magdalena, Forey Romain, Pardo Benjamin, Grzelak Marta, Nde Jules, Mitra Abhishek, Kudlicki Andrzej, Crosetto Nicola, Pasero Philippe, Rowicka Maga, Ginalski Krzysztof
Laboratory of Bioinformatics and Systems Biology, Centre of New Technologies, University of Warsaw, 02-089, Warsaw, Poland.
Department of Biochemistry and Molecular Biology, University of Texas Medical Branch at Galveston, Galveston, TX, 77555, USA.
Commun Biol. 2018 Oct 31;1:181. doi: 10.1038/s42003-018-0165-9. eCollection 2018.
Maintenance of genome stability is a key issue for cell fate that could be compromised by chromosome deletions and translocations caused by DNA double-strand breaks (DSBs). Thus development of precise and sensitive tools for DSBs labeling is of great importance for understanding mechanisms of DSB formation, their sensing and repair. Until now there has been no high resolution and specific DSB detection technique that would be applicable to any cells regardless of their size. Here, we present i-BLESS, a universal method for direct genome-wide DNA double-strand break labeling in cells immobilized in agarose beads. i-BLESS has three key advantages: it is the only unbiased method applicable to yeast, achieves a sensitivity of one break at a given position in 100,000 cells, and eliminates background noise while still allowing for fixation of samples. The method allows detection of ultra-rare breaks such as those forming spontaneously at G-quadruplexes.
维持基因组稳定性是细胞命运的关键问题，而DNA双链断裂（DSB）导致的染色体缺失和易位可能会损害这一稳定性。因此，开发用于DSB标记的精确且灵敏的工具对于理解DSB形成机制、其感应和修复至关重要。到目前为止，还没有一种高分辨率且特异性的DSB检测技术可适用于任何细胞，无论其大小如何。在此，我们展示了i-BLESS，这是一种用于在固定于琼脂糖珠中的细胞中进行全基因组范围直接DNA双链断裂标记的通用方法。i-BLESS具有三个关键优势：它是唯一适用于酵母的无偏差方法，在100,000个细胞中的给定位置实现了一个断裂的灵敏度，并且在允许固定样本的同时消除了背景噪声。该方法能够检测超罕见的断裂，例如在G-四链体处自发形成的断裂。
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