DNA Breaks and End Resection Measured Genome-wide by End Sequencing.
DNA Breaks and End Resection Measured Genome-wide by End Sequencing.
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DOI：
10.1016/j.molcel.2016.06.034
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发表时间：
2016-09-01
期刊：
影响因子：
16
通讯作者：
Nussenzweig A
中科院分区：
文献类型：
--
作者：
Canela A;Sridharan S;Sciascia N;Tubbs A;Meltzer P;Sleckman BP;Nussenzweig A
DNA双链断裂（DSB）发生在生理转录、DNA复制和抗原受体多样化过程中。DSB的错误标记或错误处理可导致病理性结构变异和突变。在这里，我们描述了一种灵敏的方法（END-seq）来监测DNA末端切除和DSB全基因组在体内的碱基对分辨率。我们利用END-seq来确定限制酶、锌指核酸酶和RAG诱导的DSB的频率和光谱。除了序列偏好，染色质特征决定了这些基因组修饰酶的所有功能。END-seq可以在10，000个不携带DSB的细胞中检测到每个细胞至少一个DSB，并且我们估计60个细胞中多达一个含有脱靶RAG切割。除了位点特异性切割，我们检测DSB分布在免疫球蛋白类转换重组过程中的扩展区域。因此，END-seq提供了全基因组DNA末端的快照，可用于理解基因组编辑特异性和染色质对DSB途径选择的影响。卡内拉等人开发了一种灵敏的定量方法，该方法在DNA修复之前提供了体内DNA双链断裂和末端切除的景观。这为更好地理解基因组不稳定性的原因和后果提供了可能性。
DNA double-strand breaks (DSBs) arise during physiological transcription, DNA replication, and antigen receptor diversification. Mistargeting or misprocessing of DSBs can result in pathological structural variation and mutation. Here we describe a sensitive method (END-seq) to monitor DNA end resection and DSBs genome-wide at base-pair resolution in vivo. We utilized END-seq to determine the frequency and spectrum of restriction-enzyme-, zinc-finger-nuclease-, and RAG-induced DSBs. Beyond sequence preference, chromatin features dictate the repertoire of these genome-modifying enzymes. END-seq can detect at least one DSB per cell among 10,000 cells not harboring DSBs, and we estimate that up to one out of 60 cells contains off-target RAG cleavage. In addition to site-specific cleavage, we detect DSBs distributed over extended regions during immunoglobulin class-switch recombination. Thus, END-seq provides a snapshot of DNA ends genome-wide, which can be utilized for understanding genome-editing specificities and the influence of chromatin on DSB pathway choice. Canela et al. develop a sensitive and quantitative method that provides a landscape of DNA double-strand breaks and end resection in vivo prior to DNA repair. This opens up the possibility for better understanding the causes and consequences of genome instability.
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影响因子：
30.8
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影响因子：
29.7
作者：
Helmink BA;Sleckman BP
通讯作者：
Sleckman BP
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14.8
作者：
Hu J;Meyers RM;Dong J;Panchakshari RA;Alt FW;Frock RL
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Barlow JH;Faryabi RB;Callén E;Wong N;Malhowski A;Chen HT;Gutierrez-Cruz G;Sun HW;McKinnon P;Wright G;Casellas R;Robbiani DF;Staudt L;Fernandez-Capetillo O;Nussenzweig A
通讯作者：
Nussenzweig A
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