High-Throughput Screening Approach for Identifying Compounds That Inhibit Nonhomologous End Joining.
High-Throughput Screening Approach for Identifying Compounds That Inhibit Nonhomologous End Joining.
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DOI：
10.1177/2472555217746324
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发表时间：
2018-08
期刊：
影响因子：
--
通讯作者：
Sleckman BP
中科院分区：
文献类型：
--
作者：
Bredemeyer AL;Edwards BS;Haynes MK;Morales AJ;Wang Y;Ursu O;Waller A;Sklar LA;Sleckman BP
DNA double strand breaks (DSBs) are repaired primarily by homologous recombination (HR) or non-homologous end joining (NHEJ). Compounds that modulate HR have shown promise as cancer therapeutics. The V(D)J recombination reaction, which assembles antigen receptor genes in lymphocytes, is initiated by the introduction of DNA DSBs at two recombining gene segments by the RAG endonuclease followed by the NHEJ-mediated repair of these DSBs. Here, using HyperCyt automated flow cytometry, we develop a robust high throughput screening (HTS) assay for NHEJ that utilizes engineered pre-B cell lines where the V(D)J recombination reaction can be induced and monitored at a single cell level. This approach, novel in processing four 384-well plates at a time in parallel, was used to screen the National Cancer Institute NeXT library to identify compounds that inhibit V(D)J recombination and NHEJ. Assessment of cell light scattering characteristics at the primary HTS stage (83,536 compounds) enabled elimination of 60% of apparent hits as false positives. Although all of the active compounds that we identified had an inhibitory effect on RAG cleavage, we have established this as an approach that could identify compounds that inhibit RAG cleavage or NHEJ using new chemical libraries.
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影响因子：
29.7
作者：
Helmink BA;Sleckman BP
通讯作者：
Sleckman BP
影响因子：
64.8
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Zha, Shan;Guo, Chunguang;Boboila, Cristian;Oksenych, Valentyn;Cheng, Hwei-Ling;Zhang, Yu;Wesemann, Duane R.;Yuen, Grace;Patel, Harin;Goff, Peter H.;Dubois, Richard L.;Alt, Frederick W.
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16
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Li, Gang;Alt, Frederick W.;Cheng, Hwei-Ling;Brush, James W.;Goff, Peter H.;Murphy, Mike M.;Franco, Sonia;Zhang, Yu;Zha, Shan
通讯作者：
Zha, Shan
影响因子：
16
作者：
Ciccia A;Elledge SJ
通讯作者：
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