van de Kooij Bert, van Attikum Haico
Department of Human Genetics, Leiden University Medical Center, Leiden, Netherlands.
Front Genet. 2022 Feb 14;12:809832. doi: 10.3389/fgene.2021.809832. eCollection 2021.
Repair of DNA Double-Strand Breaks (DSBs) can be error-free or highly mutagenic, depending on which of multiple mechanistically distinct pathways repairs the break. Hence, DSB-repair pathway choice directly affects genome integrity, and it is therefore of interest to understand the parameters that direct repair towards a specific pathway. This has been intensively studied using genomic reporter constructs, in which repair of a site-specific DSB by the pathway of interest generates a quantifiable phenotype, generally the expression of a fluorescent protein. The current developments in genome editing with targetable nucleases like Cas9 have increased reporter usage and accelerated the generation of novel reporter constructs. Considering these recent advances, this review will discuss and compare the available DSB-repair pathway reporters, provide essential considerations to guide reporter choice, and give an outlook on potential future developments.
DNA双链断裂（DSBs）的修复可以是无差错的，也可以是高度诱变的，这取决于多种机制不同的途径中的哪一种修复该断裂。因此，DSB修复途径的选择直接影响基因组完整性，所以了解指导修复走向特定途径的参数很有意义。使用基因组报告构建体对此进行了深入研究，其中感兴趣的途径对位点特异性DSB的修复产生可量化的表型，通常是荧光蛋白的表达。像Cas9这样的可靶向核酸酶在基因组编辑方面的当前进展增加了报告构建体的使用，并加速了新型报告构建体的产生。考虑到这些最新进展，本综述将讨论和比较现有的DSB修复途径报告构建体，提供指导报告构建体选择的基本注意事项，并展望潜在的未来发展。
