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New Tools to Study DNA Double-Strand Break Repair Pathway Choice

AuthorsGómez-Cabello, Daniel ; Jimeno, Sonia ; Fernández-Ávila, María Jesús ; Huertas Sánchez, Pablo
Issue Date14-Oct-2013
PublisherPublic Library of Science
CitationPLoS ONE 8(10): e77206 (2013)
AbstractA broken DNA molecule is difficult to repair, highly mutagenic, and extremely cytotoxic. Such breaks can be repaired by homology-independent or homology-directed mechanisms. Little is known about the network that controls the repair pathway choice except that a licensing step for homology-mediated repair exists, called DNA-end resection. The choice between these two repair pathways is a key event for genomic stability maintenance, and an imbalance of the ratio is directly linked with human diseases, including cancer. Here we present novel reporters to study the balance between both repair options in human cells. In these systems, a double-strand break can be alternatively repaired by homology-independent or -dependent mechanisms, leading to the accumulation of distinct fluorescent proteins. These reporters thus allow the balance between both repair pathways to be analyzed in different experimental setups. We validated the reporters by analyzing the effect of protein downregulation of the DNA end resection and non-homologous end-joining pathways. Finally, we analyzed the role of the DNA damage response on double-strand break (DSB) repair mechanism selection. Our reporters could be used in the future to understand the roles of specific factors, whole pathways, or drugs in DSB repair pathway choice, or for genome-wide screening. Moreover, our findings can be applied to increase gene-targeting efficiency, making it a beneficial tool for a broad audience in the biological sciences. © 2013 Gomez-Cabello et al.
Publisher version (URL)http://dx.doi.org/10.1371/journal.pone.0077206
Identifiersdoi: 10.1371/journal.pone.0077206
issn: 1932-6203
Appears in Collections:(CABIMER) Artículos
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