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Updated: Sep 15, 2025

Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter
Published on: March 31, 2022
A CRISPR-Cas9-based tool for dose-dependent DNA damage detection
Valentyn Oksenych1, Pavlo Petakh2, Denis Kainov1
1Department of Clinical and Molecular Medicine (IKOM), Norwegian University of Science and Technology, Trondheim, Norway.
Abstract:
Auboiron et al. developed a CRISPR-Cas9-based system in yeast Saccharomyces cerevisiae that allows precise, dose-dependent induction of DNA double-strand breaks by targeting Ty retrotransposons. This system combines sequence specificity with control over break numbers (×1, ×15, or ×59), enabling a more detailed and precise study of DNA damage response. It revealed that the key DNA damage response kinase, Tel1, localizes to the nuclear periphery and forms multiple foci after induction of DNA double-strand breaks. The system also exposed limitations in Cas9 availability at higher break levels and offers a scalable platform for studying genome stability across organisms.
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