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On-Target CRISPR/Cas9 Activity Can Cause Undesigned Large Deletion in Mouse Zygotes
Genome engineering has been tremendously affected by the appearance of the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (CRISPR/Cas9)-based approach. Initially discovered as an adaptive immune system for prokaryotes, the method has rapidly evolved ov...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7279260/ https://www.ncbi.nlm.nih.gov/pubmed/32443745 http://dx.doi.org/10.3390/ijms21103604 |
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author | Korablev, Alexey Lukyanchikova, Varvara Serova, Irina Battulin, Nariman |
author_facet | Korablev, Alexey Lukyanchikova, Varvara Serova, Irina Battulin, Nariman |
author_sort | Korablev, Alexey |
collection | PubMed |
description | Genome engineering has been tremendously affected by the appearance of the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (CRISPR/Cas9)-based approach. Initially discovered as an adaptive immune system for prokaryotes, the method has rapidly evolved over the last decade, overtaking multiple technical challenges and scientific tasks and becoming one of the most effective, reliable, and easy-to-use technologies for precise genomic manipulations. Despite its undoubtable advantages, CRISPR/Cas9 technology cannot ensure absolute accuracy and predictability of genomic editing results. One of the major concerns, especially for clinical applications, is mutations resulting from error-prone repairs of CRISPR/Cas9-induced double-strand DNA breaks. In some cases, such error-prone repairs can cause unpredicted and unplanned large genomic modifications within the CRISPR/Cas9 on-target site. Here we describe the largest, to the best of our knowledge, undesigned on-target deletion with a size of ~293 kb that occurred after the cytoplasmic injection of CRISPR/Cas9 system components into mouse zygotes and speculate about its origin. We suppose that deletion occurred as a result of the truncation of one of the ends of a double-strand break during the repair. |
format | Online Article Text |
id | pubmed-7279260 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-72792602020-06-15 On-Target CRISPR/Cas9 Activity Can Cause Undesigned Large Deletion in Mouse Zygotes Korablev, Alexey Lukyanchikova, Varvara Serova, Irina Battulin, Nariman Int J Mol Sci Article Genome engineering has been tremendously affected by the appearance of the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (CRISPR/Cas9)-based approach. Initially discovered as an adaptive immune system for prokaryotes, the method has rapidly evolved over the last decade, overtaking multiple technical challenges and scientific tasks and becoming one of the most effective, reliable, and easy-to-use technologies for precise genomic manipulations. Despite its undoubtable advantages, CRISPR/Cas9 technology cannot ensure absolute accuracy and predictability of genomic editing results. One of the major concerns, especially for clinical applications, is mutations resulting from error-prone repairs of CRISPR/Cas9-induced double-strand DNA breaks. In some cases, such error-prone repairs can cause unpredicted and unplanned large genomic modifications within the CRISPR/Cas9 on-target site. Here we describe the largest, to the best of our knowledge, undesigned on-target deletion with a size of ~293 kb that occurred after the cytoplasmic injection of CRISPR/Cas9 system components into mouse zygotes and speculate about its origin. We suppose that deletion occurred as a result of the truncation of one of the ends of a double-strand break during the repair. MDPI 2020-05-20 /pmc/articles/PMC7279260/ /pubmed/32443745 http://dx.doi.org/10.3390/ijms21103604 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Korablev, Alexey Lukyanchikova, Varvara Serova, Irina Battulin, Nariman On-Target CRISPR/Cas9 Activity Can Cause Undesigned Large Deletion in Mouse Zygotes |
title | On-Target CRISPR/Cas9 Activity Can Cause Undesigned Large Deletion in Mouse Zygotes |
title_full | On-Target CRISPR/Cas9 Activity Can Cause Undesigned Large Deletion in Mouse Zygotes |
title_fullStr | On-Target CRISPR/Cas9 Activity Can Cause Undesigned Large Deletion in Mouse Zygotes |
title_full_unstemmed | On-Target CRISPR/Cas9 Activity Can Cause Undesigned Large Deletion in Mouse Zygotes |
title_short | On-Target CRISPR/Cas9 Activity Can Cause Undesigned Large Deletion in Mouse Zygotes |
title_sort | on-target crispr/cas9 activity can cause undesigned large deletion in mouse zygotes |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7279260/ https://www.ncbi.nlm.nih.gov/pubmed/32443745 http://dx.doi.org/10.3390/ijms21103604 |
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