On-target CRISPR/CAS9 activity can cause undesigned large deletion in mouse zygotes

Alexey Korablev, Varvara Lukyanchikova, Irina Serova, Nariman Battulin

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)

Abstract

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.

Original languageEnglish
Article number3604
Number of pages14
JournalInternational Journal of Molecular Sciences
Volume21
Issue number10
DOIs
Publication statusPublished - 20 May 2020

Keywords

  • CRISPR/Cas9
  • Cytoplasmic microinjections
  • Kit knockout mice
  • Large deletion
  • On-target deletions
  • Truncation
  • Zygotic microinjections
  • CRISPR
  • VARIANTS
  • KIT GENE
  • DOMINANT-WHITE
  • large deletion
  • on-target deletions
  • GENOME
  • cytoplasmic microinjections
  • ONE-STEP GENERATION
  • DELIVERY
  • zygotic microinjections
  • CRISPR-CAS9 NUCLEASES
  • CAS9 RIBONUCLEOPROTEIN
  • MICE
  • Cas9
  • MUTATIONS
  • truncation

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