i-BLESS is an ultra-sensitive method for detection of DNA double-strand breaks.
i-BLESS is an ultra-sensitive method for detection of DNA double-strand breaks.
复制标题
DOI：
10.1038/s42003-018-0165-9
复制
发表时间：
2018
影响因子：
5.9
通讯作者：
Ginalski K
中科院分区：
文献类型：
--
作者：
Biernacka A;Zhu Y;Skrzypczak M;Forey R;Pardo B;Grzelak M;Nde J;Mitra A;Kudlicki A;Crosetto N;Pasero P;Rowicka M;Ginalski K
基因组稳定性的维持是细胞命运的关键问题，细胞命运可能会受到DNA双链断裂（DSB）引起的染色体缺失和易位的影响。因此，开发精确和灵敏的DSB标记工具对于理解DSB形成、传感和修复机制具有重要意义。到目前为止，还没有适用于任何细胞的高分辨率和特异性DSB检测技术，无论其大小如何。在这里，我们提出了i-BLESS，一个通用的方法，直接全基因组DNA双链断裂标记固定在琼脂糖珠的细胞。i-BLESS具有三个关键优势：它是适用于酵母的唯一无偏方法，在100，000个细胞中的给定位置实现一个断裂的灵敏度，并消除背景噪声，同时仍然允许固定样品。该方法允许检测超罕见的断裂，如那些自发形成的G-四链体。安娜Biernacka，Yingjie Zhu等人提出了i-BLESS，一种检测全基因组DNA双链断裂的通用方法，在此针对酵母进行了优化。通过将细胞固定在琼脂糖珠上，作者能够实现试剂的有效扩散和双链断裂的标记，包括超罕见的断裂，如G-四链体处的断裂。
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. Anna Biernacka, Yingjie Zhu et al. present i-BLESS, a universal method for detecting genome-wide DNA double strand breaks, optimized here for yeast. By immobilizing cells on agarose beads, the authors are able to achieve efficient diffusion of reagents and labeling of double strand breaks, including ultra-rare breaks such as those at G-quadruplexes.
登录
查看更多内容
影响因子：
4.5
作者：
Chung WH;Zhu Z;Papusha A;Malkova A;Ira G
通讯作者：
Ira G
影响因子：
48
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
通讯作者：
Dikic, Ivan
影响因子：
3.7
作者：
Mitra A;Skrzypczak M;Ginalski K;Rowicka M
通讯作者：
Rowicka M
影响因子：
16
作者：
Canela A;Sridharan S;Sciascia N;Tubbs A;Meltzer P;Sleckman BP;Nussenzweig A
通讯作者：
Nussenzweig A
如直接染色体分析所揭示的那样，APN1和APN2核酸内切酶可以防止在发芽的酵母中积累与修复相关的DNA断裂。
影响因子：
14.9
作者：
Ma W;Resnick MA;Gordenin DA
通讯作者：
Gordenin DA