Mutation-Independent Allele-Specific Editing by CRISPR-Cas9, a Novel Approach to Treat Autosomal Dominant Disease
Copyright © 2020 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved..
CRISPR-Cas9 provides a tool to treat autosomal dominant disease by non-homologous end joining (NHEJ) gene disruption of the mutant allele. In order to discriminate between wild-type and mutant alleles, Streptococcus pyogenes Cas9 (SpCas9) must be able to detect a single nucleotide change. Allele-specific editing can be achieved by using either a guide-specific approach, in which the missense mutation is found within the guide sequence, or a protospacer-adjacent motif (PAM)-specific approach, in which the missense mutation generates a novel PAM. While both approaches have been shown to offer allele specificity in certain contexts, in cases where numerous missense mutations are associated with a particular disease, such as TGFBI (transforming growth factor β-induced) corneal dystrophies, it is neither possible nor realistic to target each mutation individually. In this study, we demonstrate allele-specific CRISPR gene editing independent of the disease-causing mutation that is capable of achieving complete allele discrimination, and we propose it as a targeting approach for autosomal dominant disease. Our approach utilizes natural variants in the target region that contain a PAM on one allele that lies in cis with the causative mutation, removing the constraints of a mutation-dependent approach. Our innovative patient-specific guide design approach takes into account the patient's individual genetic make-up, allowing on- and off-target activity to be assessed in a personalized manner.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2020 |
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| Erschienen: |
2020 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:28 |
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| Enthalten in: |
Molecular therapy : the journal of the American Society of Gene Therapy - 28(2020), 8 vom: 05. Aug., Seite 1846-1857 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Christie, Kathleen A [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 12.07.2021 Date Revised 04.01.2024 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1016/j.ymthe.2020.05.002 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM309987091 |
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| 520 | |a Copyright © 2020 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved. | ||
| 520 | |a CRISPR-Cas9 provides a tool to treat autosomal dominant disease by non-homologous end joining (NHEJ) gene disruption of the mutant allele. In order to discriminate between wild-type and mutant alleles, Streptococcus pyogenes Cas9 (SpCas9) must be able to detect a single nucleotide change. Allele-specific editing can be achieved by using either a guide-specific approach, in which the missense mutation is found within the guide sequence, or a protospacer-adjacent motif (PAM)-specific approach, in which the missense mutation generates a novel PAM. While both approaches have been shown to offer allele specificity in certain contexts, in cases where numerous missense mutations are associated with a particular disease, such as TGFBI (transforming growth factor β-induced) corneal dystrophies, it is neither possible nor realistic to target each mutation individually. In this study, we demonstrate allele-specific CRISPR gene editing independent of the disease-causing mutation that is capable of achieving complete allele discrimination, and we propose it as a targeting approach for autosomal dominant disease. Our approach utilizes natural variants in the target region that contain a PAM on one allele that lies in cis with the causative mutation, removing the constraints of a mutation-dependent approach. Our innovative patient-specific guide design approach takes into account the patient's individual genetic make-up, allowing on- and off-target activity to be assessed in a personalized manner | ||
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| 650 | 4 | |a CRISPR-Cas9 | |
| 650 | 4 | |a allele specificity | |
| 650 | 4 | |a autosomal dominant disease | |
| 650 | 4 | |a gene therapy | |
| 650 | 4 | |a patient-specific | |
| 650 | 4 | |a personalised medicine | |
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| 700 | 1 | |a Robertson, Louise J |e verfasserin |4 aut | |
| 700 | 1 | |a Conway, Caroline |e verfasserin |4 aut | |
| 700 | 1 | |a Blighe, Kevin |e verfasserin |4 aut | |
| 700 | 1 | |a DeDionisio, Larry A |e verfasserin |4 aut | |
| 700 | 1 | |a Chao-Shern, Connie |e verfasserin |4 aut | |
| 700 | 1 | |a Kowalczyk, Amanda M |e verfasserin |4 aut | |
| 700 | 1 | |a Marshall, John |e verfasserin |4 aut | |
| 700 | 1 | |a Turnbull, Doug |e verfasserin |4 aut | |
| 700 | 1 | |a Nesbit, M Andrew |e verfasserin |4 aut | |
| 700 | 1 | |a Moore, C B Tara |e verfasserin |4 aut | |
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