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Safeguarding genome integrity during gene-editing therapy in a mouse model of age-related macular degeneration

Author

Listed:
  • Jianhang Yin

    (Peking University)

  • Kailun Fang

    (Chinese Academy of Sciences)

  • Yanxia Gao

    (Chinese Academy of Sciences)

  • Liqiong Ou

    (Peking University)

  • Shaopeng Yuan

    (Peking University)

  • Changchang Xin

    (Peking University)

  • Weiwei Wu

    (Chinese Academy of Sciences)

  • Wei-wei Wu

    (Chinese Academy of Sciences)

  • Jiaxu Hong

    (Fudan University
    the Affiliated Hospital of Guizhou Medical University)

  • Hui Yang

    (Chinese Academy of Sciences)

  • Jiazhi Hu

    (Peking University)

Abstract

Ensuring genome safety during gene editing is crucial for clinical translation of the high-efficient CRISPR-Cas9 toolbox. Therefore, the undesired events including chromosomal translocations, vector integrations, and large deletions arising during therapeutic gene editing remain to be adequately addressed or tackled in vivo. Here, we apply CRISPR-Cas9TX in comparison to CRISPR-Cas9 to target Vegfa for the treatment of age-related macular degeneration (AMD) disease in a mouse model. AAV delivery of both CRISPR-Cas9 and CRISPR-Cas9TX can efficiently inhibit laser-induced neovascularization. Importantly, Cas9TX almost eliminates chromosomal translocations that occur at a frequency of approximately 1% in Cas9-edited mouse retinal cells. Strikingly, the widely observed AAV integration at the target Vegfa site is also greatly reduced from nearly 50% of edited events to the background level during Cas9TX editing. Our findings reveal that chromosomal structural variations routinely occur during in vivo genome editing and highlight Cas9TX as a superior form of Cas9 for in vivo gene disruption.

Suggested Citation

  • Jianhang Yin & Kailun Fang & Yanxia Gao & Liqiong Ou & Shaopeng Yuan & Changchang Xin & Weiwei Wu & Wei-wei Wu & Jiaxu Hong & Hui Yang & Jiazhi Hu, 2022. "Safeguarding genome integrity during gene-editing therapy in a mouse model of age-related macular degeneration," Nature Communications, Nature, vol. 13(1), pages 1-8, December.
  • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-35640-4
    DOI: 10.1038/s41467-022-35640-4
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    References listed on IDEAS

    as
    1. Jianhang Yin & Rusen Lu & Changchang Xin & Yuhong Wang & Xinyu Ling & Dong Li & Weiwei Zhang & Mengzhu Liu & Wutao Xie & Lingyun Kong & Wen Si & Ping Wei & Bingbing Xiao & Hsiang-Ying Lee & Tao Liu & , 2022. "Cas9 exo-endonuclease eliminates chromosomal translocations during genome editing," Nature Communications, Nature, vol. 13(1), pages 1-14, December.
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    3. Eunji Kim & Taeyoung Koo & Sung Wook Park & Daesik Kim & Kyoungmi Kim & Hee-Yeon Cho & Dong Woo Song & Kyu Jun Lee & Min Hee Jung & Seokjoong Kim & Jin Hyoung Kim & Jeong Hun Kim & Jin-Soo Kim, 2017. "In vivo genome editing with a small Cas9 orthologue derived from Campylobacter jejuni," Nature Communications, Nature, vol. 8(1), pages 1-12, April.
    4. Grégoire Cullot & Julian Boutin & Jérôme Toutain & Florence Prat & Perrine Pennamen & Caroline Rooryck & Martin Teichmann & Emilie Rousseau & Isabelle Lamrissi-Garcia & Véronique Guyonnet-Duperat & Al, 2019. "CRISPR-Cas9 genome editing induces megabase-scale chromosomal truncations," Nature Communications, Nature, vol. 10(1), pages 1-14, December.
    5. Ha Youn Shin & Chaochen Wang & Hye Kyung Lee & Kyung Hyun Yoo & Xianke Zeng & Tyler Kuhns & Chul Min Yang & Teresa Mohr & Chengyu Liu & Lothar Hennighausen, 2017. "CRISPR/Cas9 targeting events cause complex deletions and insertions at 17 sites in the mouse genome," Nature Communications, Nature, vol. 8(1), pages 1-10, August.
    6. Killian S. Hanlon & Benjamin P. Kleinstiver & Sara P. Garcia & Mikołaj P. Zaborowski & Adrienn Volak & Stefan E. Spirig & Alissa Muller & Alexander A. Sousa & Shengdar Q. Tsai & Niclas E. Bengtsson & , 2019. "High levels of AAV vector integration into CRISPR-induced DNA breaks," Nature Communications, Nature, vol. 10(1), pages 1-11, December.
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    Cited by:

    1. Jianli Tao & Daniel E. Bauer & Roberto Chiarle, 2023. "Assessing and advancing the safety of CRISPR-Cas tools: from DNA to RNA editing," Nature Communications, Nature, vol. 14(1), pages 1-16, December.
    2. Milan Gautam & Antony Jozic & Grace Li-Na Su & Marco Herrera-Barrera & Allison Curtis & Sebastian Arrizabalaga & Wayne Tschetter & Renee C. Ryals & Gaurav Sahay, 2023. "Lipid nanoparticles with PEG-variant surface modifications mediate genome editing in the mouse retina," Nature Communications, Nature, vol. 14(1), pages 1-12, December.

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