Characterize compounds that affect V(D)J recombination and chromsomal translocation using MP-HTGTS

Chromosome translocation plays a crucial role in the development of tumors, but its underlying biological mechanisms, especially in human cells, remain poorly understood. In this study, we aimed to shed light on the mechanisms governing chromosome translocation in K562 cells. To investigate this, we initiated the process by inducing two double-strand breaks (DSBs) using Cas9 sgRNA and subsequently quantified the frequency of chromosome translocation events. Furthermore, we explored the influence of several compounds by introducing them alongside the DSBs, allowing us to assess their impact on chromosome translocation. Our research unveiled the identification of two noteworthy compounds, HDAC inhibitors and HSP90 inhibitors, which were found to significantly enhance the occurrence of chromosome translocation. These findings contribute to a deeper understanding of the molecular processes involved in tumorigenesis and provide potential targets for future therapeutic interventions. We induced DNA double-strand breaks (DSBs) using CRISPR-Cas9 technology in K562-derived cell line K562-Bcl2. These cell lines were subjected to different treatment conditions. To capture translocation events, we utilized one of the DSB site RAG1L as a bait, which allowed us to detect its ligation with the remaining DSB site(s) using high-throughput genome translocation sequencing (HTGTS) in microplate.