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• W2772068152 abstract "DNA double-strand breaks (DSBs) are repaired primarily by homologous recombination (HR) or nonhomologous 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 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." @default.
• W2772068152 created "2017-12-22" @default.
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• W2772068152 date "2018-08-01" @default.
• W2772068152 modified "2023-10-18" @default.
• W2772068152 title "High-Throughput Screening Approach for Identifying Compounds That Inhibit Nonhomologous End Joining" @default.
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• W2772068152 doi "https://doi.org/10.1177/2472555217746324" @default.
• W2772068152 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/5997544" @default.
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