CRISPR-Cas9 causes chromosomal instability and rearrangements in cancer cell lines, detectable by cytogenetic methods
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 t...
| Main Authors: | , , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
Mary Ann Liebert Inc.
2019
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| _version_ | 1797096943205220352 |
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| author | Rayner, E Durin, M Thomas, R Moralli, D O'Cathail, S Tomlinson, I Green, C Lewis, A |
| author_facet | Rayner, E Durin, M Thomas, R Moralli, D O'Cathail, S Tomlinson, I Green, C Lewis, A |
| author_sort | Rayner, E |
| collection | OXFORD |
| description | 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. |
| first_indexed | 2024-03-07T04:48:41Z |
| format | Journal article |
| id | oxford-uuid:d435d6ce-0167-48c8-8789-6d3e890d28b7 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T04:48:41Z |
| publishDate | 2019 |
| publisher | Mary Ann Liebert Inc. |
| record_format | dspace |
| spelling | oxford-uuid:d435d6ce-0167-48c8-8789-6d3e890d28b72022-03-27T08:16:42ZCRISPR-Cas9 causes chromosomal instability and rearrangements in cancer cell lines, detectable by cytogenetic methodsJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:d435d6ce-0167-48c8-8789-6d3e890d28b7EnglishSymplectic Elements at OxfordMary Ann Liebert Inc.2019Rayner, EDurin, MThomas, RMoralli, DO'Cathail, STomlinson, IGreen, CLewis, ACRISPR-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. |
| spellingShingle | Rayner, E Durin, M Thomas, R Moralli, D O'Cathail, S Tomlinson, I Green, C Lewis, A CRISPR-Cas9 causes chromosomal instability and rearrangements in cancer cell lines, detectable by cytogenetic methods |
| title | CRISPR-Cas9 causes chromosomal instability and rearrangements in cancer cell lines, detectable by cytogenetic methods |
| title_full | CRISPR-Cas9 causes chromosomal instability and rearrangements in cancer cell lines, detectable by cytogenetic methods |
| title_fullStr | CRISPR-Cas9 causes chromosomal instability and rearrangements in cancer cell lines, detectable by cytogenetic methods |
| title_full_unstemmed | CRISPR-Cas9 causes chromosomal instability and rearrangements in cancer cell lines, detectable by cytogenetic methods |
| title_short | CRISPR-Cas9 causes chromosomal instability and rearrangements in cancer cell lines, detectable by cytogenetic methods |
| title_sort | crispr cas9 causes chromosomal instability and rearrangements in cancer cell lines detectable by cytogenetic methods |
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