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Low-dose AAV-CRISPR-mediated liver-specific knock-in restored hemostasis in neonatal hemophilia B mice with subtle antibody response
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 ho...
Autores principales: | , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9700734/ https://www.ncbi.nlm.nih.gov/pubmed/36434000 http://dx.doi.org/10.1038/s41467-022-34898-y |
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author | He, Xiangjun Zhang, Zhenjie Xue, Junyi Wang, Yaofeng Zhang, Siqi Wei, Junkang Zhang, Chenzi Wang, Jue Urip, Brian Anugerah Ngan, Chun Christopher Sun, Junjiang Li, Yuefeng Lu, Zhiqian Zhao, Hui Pei, Duanqing Li, Chi-Kong Feng, Bo |
author_facet | He, Xiangjun Zhang, Zhenjie Xue, Junyi Wang, Yaofeng Zhang, Siqi Wei, Junkang Zhang, Chenzi Wang, Jue Urip, Brian Anugerah Ngan, Chun Christopher Sun, Junjiang Li, Yuefeng Lu, Zhiqian Zhao, Hui Pei, Duanqing Li, Chi-Kong Feng, Bo |
author_sort | He, Xiangjun |
collection | PubMed |
description | 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 × 10(9) vg/neonate and 1 × 10(10) vg/adult mouse) through liver-specific gene knock-in using hyperactive hF9(R338L) 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. |
format | Online Article Text |
id | pubmed-9700734 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-97007342022-11-27 Low-dose AAV-CRISPR-mediated liver-specific knock-in restored hemostasis in neonatal hemophilia B mice with subtle antibody response He, Xiangjun Zhang, Zhenjie Xue, Junyi Wang, Yaofeng Zhang, Siqi Wei, Junkang Zhang, Chenzi Wang, Jue Urip, Brian Anugerah Ngan, Chun Christopher Sun, Junjiang Li, Yuefeng Lu, Zhiqian Zhao, Hui Pei, Duanqing Li, Chi-Kong Feng, Bo Nat Commun Article 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 × 10(9) vg/neonate and 1 × 10(10) vg/adult mouse) through liver-specific gene knock-in using hyperactive hF9(R338L) 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. Nature Publishing Group UK 2022-11-25 /pmc/articles/PMC9700734/ /pubmed/36434000 http://dx.doi.org/10.1038/s41467-022-34898-y Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article He, Xiangjun Zhang, Zhenjie Xue, Junyi Wang, Yaofeng Zhang, Siqi Wei, Junkang Zhang, Chenzi Wang, Jue Urip, Brian Anugerah Ngan, Chun Christopher Sun, Junjiang Li, Yuefeng Lu, Zhiqian Zhao, Hui Pei, Duanqing Li, Chi-Kong Feng, Bo Low-dose AAV-CRISPR-mediated liver-specific knock-in restored hemostasis in neonatal hemophilia B mice with subtle antibody response |
title | Low-dose AAV-CRISPR-mediated liver-specific knock-in restored hemostasis in neonatal hemophilia B mice with subtle antibody response |
title_full | Low-dose AAV-CRISPR-mediated liver-specific knock-in restored hemostasis in neonatal hemophilia B mice with subtle antibody response |
title_fullStr | Low-dose AAV-CRISPR-mediated liver-specific knock-in restored hemostasis in neonatal hemophilia B mice with subtle antibody response |
title_full_unstemmed | Low-dose AAV-CRISPR-mediated liver-specific knock-in restored hemostasis in neonatal hemophilia B mice with subtle antibody response |
title_short | Low-dose AAV-CRISPR-mediated liver-specific knock-in restored hemostasis in neonatal hemophilia B mice with subtle antibody response |
title_sort | low-dose aav-crispr-mediated liver-specific knock-in restored hemostasis in neonatal hemophilia b mice with subtle antibody response |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9700734/ https://www.ncbi.nlm.nih.gov/pubmed/36434000 http://dx.doi.org/10.1038/s41467-022-34898-y |
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