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Monitoring Double-Strand Break Repair of Trinucleotide Repeats Using a Yeast Fluorescent Reporter Assay

Abstract : Cells can repair a double-strand break (DSB) by homologous recombination if a homologous sequence is provided as a template. This can be achieved by classical gene conversion (with or without crossover) or by single-strand annealing (SSA) between two direct repeat sequences flanking the DSB. To initiate SSA, single-stranded regions are needed adjacent to the break, extending up to the direct repeats in such a way that complementary strands can anneal to each other to repair the DSB. In the present protocol, we describe a GFP reporter assay in Saccharomyces cerevisiae allowing for the quantification of nuclease efficacy at inducing a DSB, by monitoring the reconstitution of a functional GFP gene whose expression can be rapidly quantified by flow cytometry.
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https://hal-pasteur.archives-ouvertes.fr/pasteur-02864616
Contributor : Guy-Franck Richard <>
Submitted on : Thursday, June 11, 2020 - 11:00:39 AM
Last modification on : Saturday, October 17, 2020 - 3:14:04 AM

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Lucie Poggi, Bruno Dumas, Guy-Franck Richard. Monitoring Double-Strand Break Repair of Trinucleotide Repeats Using a Yeast Fluorescent Reporter Assay. Guy-Franck Richard. Trinucleotide Repeats : Methods and Protocols, Springer Science; Business Media, LLC, pp.113-120, 2019, 978-1-4939-9783-1. ⟨10.1007/978-1-4939-9784-8_7⟩. ⟨pasteur-02864616⟩

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