論文

査読有り 国際誌
2020年3月13日

Extracellular nanovesicles for packaging of CRISPR-Cas9 protein and sgRNA to induce therapeutic exon skipping.

Nature communications
  • Peter Gee
  • Mandy S Y Lung
  • Yuya Okuzaki
  • Noriko Sasakawa
  • Takahiro Iguchi
  • Yukimasa Makita
  • Hiroyuki Hozumi
  • Yasutomo Miura
  • Lucy F Yang
  • Mio Iwasaki
  • Xiou H Wang
  • Matthew A Waller
  • Nanako Shirai
  • Yasuko O Abe
  • Yoko Fujita
  • Kei Watanabe
  • Akihiro Kagita
  • Kumiko A Iwabuchi
  • Masahiko Yasuda
  • Huaigeng Xu
  • Takeshi Noda
  • Jun Komano
  • Hidetoshi Sakurai
  • Naoto Inukai
  • Akitsu Hotta
  • 全て表示

11
1
開始ページ
1334
終了ページ
1334
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1038/s41467-020-14957-y

Prolonged expression of the CRISPR-Cas9 nuclease and gRNA from viral vectors may cause off-target mutagenesis and immunogenicity. Thus, a transient delivery system is needed for therapeutic genome editing applications. Here, we develop an extracellular nanovesicle-based ribonucleoprotein delivery system named NanoMEDIC by utilizing two distinct homing mechanisms. Chemical induced dimerization recruits Cas9 protein into extracellular nanovesicles, and then a viral RNA packaging signal and two self-cleaving riboswitches tether and release sgRNA into nanovesicles. We demonstrate efficient genome editing in various hard-to-transfect cell types, including human induced pluripotent stem (iPS) cells, neurons, and myoblasts. NanoMEDIC also achieves over 90% exon skipping efficiencies in skeletal muscle cells derived from Duchenne muscular dystrophy (DMD) patient iPS cells. Finally, single intramuscular injection of NanoMEDIC induces permanent genomic exon skipping in a luciferase reporter mouse and in mdx mice, indicating its utility for in vivo genome editing therapy of DMD and beyond.

リンク情報
DOI
https://doi.org/10.1038/s41467-020-14957-y
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/32170079
PubMed Central
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7070030
ID情報
  • DOI : 10.1038/s41467-020-14957-y
  • PubMed ID : 32170079
  • PubMed Central 記事ID : PMC7070030

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