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Low-dose AAV-CRISPR-mediated liver-specific knock-in restored hemostasis in neonatal hemophilia B mice with subtle antibody response

Author

Listed:
  • Xiangjun He

    (The Chinese University of Hong Kong)

  • Zhenjie Zhang

    (The Chinese University of Hong Kong)

  • Junyi Xue

    (The Chinese University of Hong Kong)

  • Yaofeng Wang

    (Chinese Academy of Sciences
    Chinese Academy of Sciences)

  • Siqi Zhang

    (The Chinese University of Hong Kong)

  • Junkang Wei

    (The Chinese University of Hong Kong)

  • Chenzi Zhang

    (The Chinese University of Hong Kong
    Chinese Academy of Sciences)

  • Jue Wang

    (The Chinese University of Hong Kong)

  • Brian Anugerah Urip

    (The Chinese University of Hong Kong)

  • Chun Christopher Ngan

    (The Chinese University of Hong Kong
    Chinese Academy of Sciences)

  • Junjiang Sun

    (University of North Carolina)

  • Yuefeng Li

    (Guangdong Landau Biotechnology Co.Ltd)

  • Zhiqian Lu

    (Shanghai Jiao Tong University Affiliated Sixth People’s Hospital)

  • Hui Zhao

    (The Chinese University of Hong Kong
    The Chinese University of Hong Kong, Shenzhen Research Institute)

  • Duanqing Pei

    (Chinese Academy of Sciences
    Westlake University)

  • Chi-Kong Li

    (The Chinese University of Hong Kong)

  • Bo Feng

    (The Chinese University of Hong Kong
    Chinese Academy of Sciences
    The Chinese University of Hong Kong, Shenzhen Research Institute)

Abstract

AAV-delivered CRISPR/Cas9 (AAV-CRISPR) has shown promising potentials in preclinical models to efficiently insert therapeutic gene sequences in somatic tissues. However, the AAV input doses required were prohibitively high and posed serious risk of toxicity. Here, we performed AAV-CRISPR mediated homology-independent knock-in at a new target site in mAlb 3’UTR and demonstrated that single dose of AAVs enabled long-term integration and expression of hF9 transgene in both adult and neonatal hemophilia B mice (mF9 −/−), yielding high levels of circulating human Factor IX (hFIX) and stable hemostasis restoration during entire 48-week observation period. Furthermore, we achieved hemostasis correction with a significantly lower AAV dose (2 × 109 vg/neonate and 1 × 1010 vg/adult mouse) through liver-specific gene knock-in using hyperactive hF9R338L variant. The plasma antibodies against Cas9 and AAV in the neonatal mice receiving low-dose AAV-CRISPR were negligible, which lent support to the development of AAV-CRISPR mediated somatic knock-in for treating inherited diseases.

Suggested Citation

  • Xiangjun He & Zhenjie Zhang & Junyi Xue & Yaofeng Wang & Siqi Zhang & Junkang Wei & Chenzi Zhang & Jue Wang & Brian Anugerah Urip & Chun Christopher Ngan & Junjiang Sun & Yuefeng Li & Zhiqian Lu & Hui, 2022. "Low-dose AAV-CRISPR-mediated liver-specific knock-in restored hemostasis in neonatal hemophilia B mice with subtle antibody response," Nature Communications, Nature, vol. 13(1), pages 1-17, December.
  • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-34898-y
    DOI: 10.1038/s41467-022-34898-y
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