CRISPR-Cas9 causes chromosomal instability and rearrangements in cancer cell lines, detectable by cytogenetic methods

Rayner, E., Durin, M.-A., Thomas, R., Moralli, D., O'Cathail, S. M. , Tomlinson, I., Green, C. M. and Lewis, A. (2019) CRISPR-Cas9 causes chromosomal instability and rearrangements in cancer cell lines, detectable by cytogenetic methods. CRISPR Journal, 2(6), pp. 406-416. (doi: 10.1089/crispr.2019.0006) (PMID:31742432) (PMCID:PMC6919265)

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Abstract

CRISPR-Cas9 has quickly become the method of choice for genome editing, with multiple publications describing technical advances and novel applications. It has been widely adopted as a tool for basic research and has significant translational and clinical potential. However, its usage has outpaced the establishment of essential and rigorous controls for unwanted off-target effects, manifested as small mutations, large deletions of target loci, or large-scale chromosomal rearrangements. A common application of CRISPR-Cas9 is as a tool for creating isogenic cell-line models to study the effects of precise mutations, or variants, on disease traits. Here, we describe the effect of standard CRISPR-Cas9 mutagenesis protocols on well characterized cancer cell lines. We demonstrate that commonly used methods for detecting correctly mutated clones fail to uncover large-scale rearrangements. We show that simple cytogenetic methods can be used to identify clones carrying chromosomal abnormalities and large mutations at target loci. These methods are quick and cost-efficient, and we suggest that such controls should be performed prior to publication of studies based on novel CRISPR-Cas9 mutated cancer cell lines.

Item Type:Articles
Additional Information:Funding was provided by a Medical Research Council New Investigator Research (MR/P000738/1). Core funding to the Wellcome Centre for Human Genetics was provided by the Wellcome Trust (090532/Z/09/Z).
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:O'Cathail, Dr Sean
Authors: Rayner, E., Durin, M.-A., Thomas, R., Moralli, D., O'Cathail, S. M., Tomlinson, I., Green, C. M., and Lewis, A.
College/School:College of Medical Veterinary and Life Sciences > Institute of Cancer Sciences
Journal Name:CRISPR Journal
Publisher:Mary Ann Liebert
ISSN:2573-1599
ISSN (Online):2573-1602
Published Online:19 November 2019
Copyright Holders:Copyright © 2019 Emily Rayner et al.
First Published:First published in CRISPR Journal 2(6): 406-416
Publisher Policy:Reproduced under a Creative Commons License

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