Precision digital mapping of endogenous and induced genomic DNA breaks by INDUCE-seq.
Precision digital mapping of endogenous and induced genomic DNA breaks by INDUCE-seq.
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DOI：
10.1038/s41467-022-31702-9
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发表时间：
2022-07-09
影响因子：
16.6
通讯作者：
中科院分区：
文献类型：
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了解断裂如何在基因组中形成和修复取决于DNA双链断裂（DSBs）的频率和位置的精确测量。这对于识别化学物质的DNA损伤潜力以及安全开发包括基因组编辑技术在内的治疗方法至关重要。目前的DSB测序方法存在高背景水平、无法准确测量低频内源性断裂和测序成本高的问题。在这里，我们描述了克服这些问题的诱导-seq，它可以同时检测由生理过程引起的低水平内源性dsb的存在，以及由限制性内切酶或CRISPR-Cas核酸酶诱导的高水平复发断裂。诱导-seq利用创新的NGS流动池富集方法，允许数字检测断裂。因此，它可以用于确定DSB修复的机制，并促进治疗性基因组编辑的安全开发。我们进一步讨论了该方法如何适用于检测其他基因组特征。了解DNA双链断裂（DSBs）如何在基因组中形成和修复取决于它们的精确测量。在这里，作者描述了诱导-序列；一种dsb检测方法，同时测量整个基因组的生理和诱导断裂。
Understanding how breaks form and are repaired in the genome depends on the accurate measurement of the frequency and position of DNA double strand breaks (DSBs). This is crucial for identification of a chemical’s DNA damage potential and for safe development of therapies, including genome editing technologies. Current DSB sequencing methods suffer from high background levels, the inability to accurately measure low frequency endogenous breaks and high sequencing costs. Here we describe INDUCE-seq, which overcomes these problems, detecting simultaneously the presence of low-level endogenous DSBs caused by physiological processes, and higher-level recurrent breaks induced by restriction enzymes or CRISPR-Cas nucleases. INDUCE-seq exploits an innovative NGS flow cell enrichment method, permitting the digital detection of breaks. It can therefore be used to determine the mechanism of DSB repair and to facilitate safe development of therapeutic genome editing. We further discuss how the method can be adapted to detect other genomic features. Understanding how DNA double strand breaks (DSBs) form and are repaired in the genome depends on their accurate measurement. Here the authors describe INDUCE-seq; a DSB-detection method that simultaneously measures physiological and induced breaks throughout the genome.
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DOI：
10.1093/bioinformatics/btq033
发表时间：
2010-03-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Quinlan AR;Hall IM
通讯作者：
Hall IM
影响因子：
5.9
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
通讯作者：
Ginalski K
影响因子：
46.9
作者：
Tsai, Shengdar Q.;Zheng, Zongli;Nguyen, Nhu T.;Liebers, Matthew;Topkar, Ved V.;Thapar, Vishal;Wyvekens, Nicolas;Khayter, Cyd;Iafrate, A. John;Le, Long P.;Aryee, Martin J.;Joung, J. Keith
通讯作者：
Joung, J. Keith
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：
56.9
作者：
Jinek, Martin;Chylinski, Krzysztof;Charpentier, Emmanuelle
通讯作者：
Charpentier, Emmanuelle