Papers

International journal
2024

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

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

Volume
19
Number
5
First page
e0288578
Last page
Language
English
Publishing type
Research paper (scientific journal)
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.

Link information
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 information
  • DOI : 10.1371/journal.pone.0288578
  • Pubmed ID : 38739603
  • Pubmed Central ID : PMC11090291

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