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Open AccessArticle

On-Target CRISPR/Cas9 Activity Can Cause Undesigned Large Deletion in Mouse Zygotes

1
Institute of Cytology and Genetics SB RAS, Novosibirsk 630090, Russia
2
Laboratory of structural and functional genome organization, Novosibirsk State University, Novosibirsk 630090, Russia
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2020, 21(10), 3604; https://doi.org/10.3390/ijms21103604
Received: 20 April 2020 / Revised: 15 May 2020 / Accepted: 19 May 2020 / Published: 20 May 2020
(This article belongs to the Section Molecular Pathology, Diagnostics, and Therapeutics)
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. View Full-Text
Keywords: CRISPR/Cas9; on-target deletions; large deletion; truncation; cytoplasmic microinjections; zygotic microinjections; Kit knockout mice CRISPR/Cas9; on-target deletions; large deletion; truncation; cytoplasmic microinjections; zygotic microinjections; Kit knockout mice
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Korablev, A.; Lukyanchikova, V.; Serova, I.; Battulin, N. On-Target CRISPR/Cas9 Activity Can Cause Undesigned Large Deletion in Mouse Zygotes. Int. J. Mol. Sci. 2020, 21, 3604.

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