Assessment and Mitigation of CRISPR‐Cas9‐Induced Nontargeted Translocations
Abstract The performance of CRISPR‐mediated genome editing near inverted repeats (IRs) potentially results in chromosomal translocations and other catastrophic rearrangements. However, the extent of this risk may be significantly underestimated because current reporter systems focus solely on site‐s...
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| Format: | Article |
| Language: | English |
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Wiley
2025-06-01
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| Series: | Advanced Science |
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| Online Access: | https://doi.org/10.1002/advs.202414415 |
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| author | Zhiyang Hou Qiyi Yi Mengying Wu Lijun Wu Fanghua Li Ting Wang Po Bian |
| author_facet | Zhiyang Hou Qiyi Yi Mengying Wu Lijun Wu Fanghua Li Ting Wang Po Bian |
| author_sort | Zhiyang Hou |
| collection | DOAJ |
| description | Abstract The performance of CRISPR‐mediated genome editing near inverted repeats (IRs) potentially results in chromosomal translocations and other catastrophic rearrangements. However, the extent of this risk may be significantly underestimated because current reporter systems focus solely on site‐specific translocations. Here, trans‐acting reporter systems in Escherichia coli are developed to detect nontargeted translocations. Markedly increased frequency of translocations following CRISPR‐Cas9 activation is observed, with the magnitude determined primarily by the length of the IRs and the proximity between Cas9 target sites and IRs. These translocations arise through a combination of intramolecular single‐strand annealing and alternative end‐joining mechanisms. Furthermore, it is discovered that introducing segments homologous to IR loci can substantially mitigate nontargeted translocations without significantly compromising CRISPR‐Cas9‐mediated editing. The study provides valuable insights into the genetic risks associated with CRISPR technologies and suggests a viable strategy for developing genetically safer CRISPR systems. |
| format | Article |
| id | doaj-art-3761b5e4f27646e9aa286bf4e72f7ebd |
| institution | OA Journals |
| issn | 2198-3844 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Wiley |
| record_format | Article |
| series | Advanced Science |
| spelling | doaj-art-3761b5e4f27646e9aa286bf4e72f7ebd2025-08-20T02:02:58ZengWileyAdvanced Science2198-38442025-06-011221n/an/a10.1002/advs.202414415Assessment and Mitigation of CRISPR‐Cas9‐Induced Nontargeted TranslocationsZhiyang Hou0Qiyi Yi1Mengying Wu2Lijun Wu3Fanghua Li4Ting Wang5Po Bian6School of Basic Medical Sciences Anhui Medical University Hefei 230032 ChinaSchool of Basic Medical Sciences Anhui Medical University Hefei 230032 ChinaSchool of Basic Medical Sciences Anhui Medical University Hefei 230032 ChinaInstitute of Physical Science and Information Technology Anhui University Hefei 230601 ChinaDepartment of Particle Therapy West German Proton Therapy Centre Essen (WPE) West German Cancer Center (WTZ) German Cancer Consortium (DKTK) University Hospital Essen 45147 Essen GermanySchool of Basic Medical Sciences Anhui Medical University Hefei 230032 ChinaSchool of Basic Medical Sciences Anhui Medical University Hefei 230032 ChinaAbstract The performance of CRISPR‐mediated genome editing near inverted repeats (IRs) potentially results in chromosomal translocations and other catastrophic rearrangements. However, the extent of this risk may be significantly underestimated because current reporter systems focus solely on site‐specific translocations. Here, trans‐acting reporter systems in Escherichia coli are developed to detect nontargeted translocations. Markedly increased frequency of translocations following CRISPR‐Cas9 activation is observed, with the magnitude determined primarily by the length of the IRs and the proximity between Cas9 target sites and IRs. These translocations arise through a combination of intramolecular single‐strand annealing and alternative end‐joining mechanisms. Furthermore, it is discovered that introducing segments homologous to IR loci can substantially mitigate nontargeted translocations without significantly compromising CRISPR‐Cas9‐mediated editing. The study provides valuable insights into the genetic risks associated with CRISPR technologies and suggests a viable strategy for developing genetically safer CRISPR systems.https://doi.org/10.1002/advs.202414415alternative end‐joiningchromosomal rearrangementCRISPR‐mediated genome editingDNA single‐strand annealingnontargeted translocation |
| spellingShingle | Zhiyang Hou Qiyi Yi Mengying Wu Lijun Wu Fanghua Li Ting Wang Po Bian Assessment and Mitigation of CRISPR‐Cas9‐Induced Nontargeted Translocations Advanced Science alternative end‐joining chromosomal rearrangement CRISPR‐mediated genome editing DNA single‐strand annealing nontargeted translocation |
| title | Assessment and Mitigation of CRISPR‐Cas9‐Induced Nontargeted Translocations |
| title_full | Assessment and Mitigation of CRISPR‐Cas9‐Induced Nontargeted Translocations |
| title_fullStr | Assessment and Mitigation of CRISPR‐Cas9‐Induced Nontargeted Translocations |
| title_full_unstemmed | Assessment and Mitigation of CRISPR‐Cas9‐Induced Nontargeted Translocations |
| title_short | Assessment and Mitigation of CRISPR‐Cas9‐Induced Nontargeted Translocations |
| title_sort | assessment and mitigation of crispr cas9 induced nontargeted translocations |
| topic | alternative end‐joining chromosomal rearrangement CRISPR‐mediated genome editing DNA single‐strand annealing nontargeted translocation |
| url | https://doi.org/10.1002/advs.202414415 |
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