Genomic Reporter Constructs to Monitor Pathway-Specific Repair of DNA Double-Strand Breaks

Front Genet. 2022 Feb 14:12:809832. doi: 10.3389/fgene.2021.809832. eCollection 2021.

Abstract

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.

Keywords: double-strand break repair pathway choice; end-joining; genomic reporter constructs; homologous recombination; single-strand annealing.

Publication types

  • Review