論文

国際誌
2018年8月16日

Biased genome editing using the local accumulation of DSB repair molecules system.

Nature communications
  • Shota Nakade
  • ,
  • Keiji Mochida
  • ,
  • Atsushi Kunii
  • ,
  • Kazuki Nakamae
  • ,
  • Tomomi Aida
  • ,
  • Kohichi Tanaka
  • ,
  • Naoaki Sakamoto
  • ,
  • Tetsushi Sakuma
  • ,
  • Takashi Yamamoto

9
1
開始ページ
3270
終了ページ
3270
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1038/s41467-018-05773-6

Selective genome editing such as gene knock-in has recently been achieved by administration of chemical enhancer or inhibitor of particular DNA double-strand break (DSB) repair pathways, as well as overexpression of pathway-specific genes. In this study, we attempt to enhance the efficiency further to secure robust gene knock-ins, by using the local accumulation of DSB repair molecules (LoAD) system. We identify CtIP as a strong enhancer of microhomology-mediated end-joining (MMEJ) repair by genetic screening, and show the knock-in-enhancing effect of CtIP LoADing. Next-generation sequencing reveals that CtIP LoADing highly increases the frequency of MMEJ-mediated integration. Selection-free, simultaneous triple gene knock-ins are also achieved with the CtIP-LoADing strategy. Moreover, by replacing the LoADing molecules and targeting strategies, this system can be applied for other specific genome engineering purposes, such as introducing longer deletions for gene disruption, independently introducing multiple mutations without chromosomal deletion, and efficiently incorporating a single-stranded oligodeoxynucleotide donor.

リンク情報
DOI
https://doi.org/10.1038/s41467-018-05773-6
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/30115916
PubMed Central
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6095859
ID情報
  • DOI : 10.1038/s41467-018-05773-6
  • PubMed ID : 30115916
  • PubMed Central 記事ID : PMC6095859

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