Genetic evaluation of CRISPR-Cas9 off-target effects from deleterious mutations on Drosophila male single X chromosome

Bian, Wei and Mcquarrie, David W. J. and Haussmann, Irmgard U. and Arnold, Roland and Soller, Matthias (2025) Genetic evaluation of CRISPR-Cas9 off-target effects from deleterious mutations on Drosophila male single X chromosome. Functional & Integrative Genomics, 25 (1). ISSN 1438-793X

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Abstract

Clustered regularly interspaced short palindromic repeats (CRISPR)-associated nuclease protein 9 (Cas9) is a powerful tool used for genome engineering, but concerns remain about off-target effects. Here we evaluate potential deleterious effects of CRISPR-Cas9 by combining sequence analysis and the genetics of the male X chromosome in a Drosophila model. Since males have only one X chromosome deleterious mutations on the X chromosome will manifest in reducing viability or result in visible phenotypes and thus provide sensitive readouts of off-target activity. Our data do not support large scale off-target effects in Drosophila . To optimize sgRNA selection, we incorporated off-target evaluation into the PlatinumCRISPr sgRNA selection tool for a broad range of organisms.

Item Type:	Article
Identification Number:	10.1007/s10142-025-01775-y
Dates:	Date Event 14 November 2025 Accepted 10 December 2025 Published Online
Uncontrolled Keywords:	CRISPR, Cas9, SgRNA, Off-target effect, Drosophila
Subjects:	CAH03 - biological and sport sciences > CAH03-01 - biosciences > CAH03-01-02 - biology (non-specific)
Divisions:	Life and Health Sciences > Life and Sports Sciences
Depositing User:	Gemma Tonks
Date Deposited:	30 Jun 2026 09:47
Last Modified:	30 Jun 2026 09:47
URI:	https://www.open-access.bcu.ac.uk/id/eprint/17088

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