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)

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 > School 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