論文

国際誌
2024年

Fusion of histone variants to Cas9 suppresses non-homologous end joining.

PloS one
  • Tomoko Kato-Inui
  • ,
  • Gou Takahashi
  • ,
  • Terumi Ono
  • ,
  • Yuichiro Miyaoka

19
5
開始ページ
e0288578
終了ページ
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1371/journal.pone.0288578

As a versatile genome editing tool, the CRISPR-Cas9 system induces DNA double-strand breaks at targeted sites to activate mainly two DNA repair pathways: HDR which allows precise editing via recombination with a homologous template DNA, and NHEJ which connects two ends of the broken DNA, which is often accompanied by random insertions and deletions. Therefore, how to enhance HDR while suppressing NHEJ is a key to successful applications that require precise genome editing. Histones are small proteins with a lot of basic amino acids that generate electrostatic affinity to DNA. Since H2A.X is involved in DNA repair processes, we fused H2A.X to Cas9 and found that this fusion protein could improve the HDR/NHEJ ratio by suppressing NHEJ. As various post-translational modifications of H2A.X play roles in the regulation of DNA repair, we also fused H2A.X mimicry variants to replicate these post-translational modifications including phosphorylation, methylation, and acetylation. However, none of them were effective to improve the HDR/NHEJ ratio. We further fused other histone variants to Cas9 and found that H2A.1 suppressed NHEJ better than H2A.X. Thus, the fusion of histone variants to Cas9 is a promising option to enhance precise genome editing.

リンク情報
DOI
https://doi.org/10.1371/journal.pone.0288578
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/38739603
PubMed Central
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11090291
ID情報
  • DOI : 10.1371/journal.pone.0288578
  • PubMed ID : 38739603
  • PubMed Central 記事ID : PMC11090291

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