TY - JOUR
T1 - Single AAV-mediated mutation replacement genome editing in limited number of photoreceptors restores vision in mice
AU - Nishiguchi, Koji M.
AU - Fujita, Kosuke
AU - Miya, Fuyuki
AU - Katayama, Shota
AU - Nakazawa, Toru
N1 - Funding Information:
We thank Ms Misane Uchiike for her help with the experiments and Professor Carlo Rivolta for the critical comments on the manuscript. This work was supported in part by the Japan Agency for Medical Research and Development (KMN, 18ek0109213h0002). The manuscript was edited by a professional English editing service (Mr. Tim Hilts). We thank the Biomedical Research Core of Tohoku University Graduate School of Medicine for technical support.
Funding Information:
K.M.N., K.F. and T.N. are listed as inventors in a patent application related to this work. The other authors declare no competing interests. The Departments of Advanced Ophthalmic Medicine and Ophthalmic Imaging and Information Analytics are endowed departments, supported by an unrestricted grant from Senju Pharmaceutical Co. (Osaka, Japan) and Topcon Co. Ltd. (Tokyo, Japan), respectively. These funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the paper.
Publisher Copyright:
© 2020, The Author(s).
PY - 2020/12/1
Y1 - 2020/12/1
N2 - Supplementing wildtype copies of functionally defective genes with adeno-associated virus (AAV) is a strategy being explored clinically for various retinal dystrophies. However, the low cargo limit of this vector allows its use in only a fraction of patients with mutations in relatively small pathogenic genes. To overcome this issue, we developed a single AAV platform that allows local replacement of a mutated sequence with its wildtype counterpart, based on combined CRISPR-Cas9 and micro-homology-mediated end-joining (MMEJ). In blind mice, the mutation replacement rescued approximately 10% of photoreceptors, resulting in an improvement in light sensitivity and an increase in visual acuity. These effects were comparable to restoration mediated by gene supplementation, which targets a greater number of photoreceptors. This strategy may be applied for the treatment of inherited disorders caused by mutations in larger genes, for which conventional gene supplementation therapy is not currently feasible.
AB - Supplementing wildtype copies of functionally defective genes with adeno-associated virus (AAV) is a strategy being explored clinically for various retinal dystrophies. However, the low cargo limit of this vector allows its use in only a fraction of patients with mutations in relatively small pathogenic genes. To overcome this issue, we developed a single AAV platform that allows local replacement of a mutated sequence with its wildtype counterpart, based on combined CRISPR-Cas9 and micro-homology-mediated end-joining (MMEJ). In blind mice, the mutation replacement rescued approximately 10% of photoreceptors, resulting in an improvement in light sensitivity and an increase in visual acuity. These effects were comparable to restoration mediated by gene supplementation, which targets a greater number of photoreceptors. This strategy may be applied for the treatment of inherited disorders caused by mutations in larger genes, for which conventional gene supplementation therapy is not currently feasible.
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U2 - 10.1038/s41467-019-14181-3
DO - 10.1038/s41467-019-14181-3
M3 - Article
C2 - 31980606
AN - SCOPUS:85078155735
VL - 11
JO - Nature Communications
JF - Nature Communications
SN - 2041-1723
IS - 1
M1 - 482
ER -