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Correction of a Genetic Disease in Mouse via Use of CRISPR-Cas9
Open accessAbstract
The CRISPR-Cas9 system has been employed to generate mutant alleles in a range of different organisms. However, so far there have not been reports of use of this system for efficient correction of a genetic disease. Here we show that mice with a dominant mutation in Crygc gene that causes cataracts could be rescued by coinjection into zygotes of Cas9 mRNA and a single-guide RNA (sgRNA) targeting the mutant allele. Correction occurred via homology-directed repair (HDR) based on an exogenously supplied oligonucleotide or the endogenous WT allele, with only rare evidence of off-target modifications. The resulting mice were fertile and able to transmit the corrected allele to their progeny. Thus, our study provides proof of principle for use of the CRISPR-Cas9 system to correct genetic disease.
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In the citing paper
“Due to the low rate of HDR recombination [109]”
From CRISPR-Cas: Converting A Bacterial Defence Mechanism into A State-of-the-Art Genetic Manipulation Tool (2019) · cites this paper as [109]
In the citing paper
“Several groups have also demonstrated in vitro correction of genetic disease genes including CFTR (cystic fibrosis)9, Crygc (cataracts)10 and DMD (Duchenne muscular dystrophy)11 in organoids or mouse zygotes.”
From Therapeutic genome editing by combined viral and non-viral delivery of CRISPR system components in vivo (2016) · cites this paper as [10]
In the citing paper
“Furthermore, CRISPR/Cas9 system holds tremendous promise for gene therapy [46–51]. It can correct causal mutations in the original genome and rescue the disease phenotypes of monogenic disorders permanently, which currently represents the most translatable field in CRISPR/Cas9-mediated disease therapy.”
From Non-viral delivery systems for CRISPR/Cas9-based genome editing: challenges and opportunities (2018) · cites this paper as [47]
In the citing paper
“CRISPR-Cas9 genome editing has been applied to correct disease-causing mutations in mouse zygotes and human cell lines for cataract5 and cystic fibrosis6”
From Genome editing with Cas9 in adult mice corrects a disease mutation and phenotype (2014) · cites this paper as [5]
In the citing paper
“In such a mouse model, germline-manipulation with CRISPR-Cas9 system was capable of correcting both the mutant gene and cataract phenotype in part of offsprings.”
From CRISPR-Cas9: a new and promising player in gene therapy (2015) · cites this paper as [30]
In the citing paper
“Interestingly, however, of the six offsprings undergoing NHEJ, two were also free of cataract thanks to coincidental frameshift restoration resulting from NHEJ-induced indel mutations, while the rest four remained cataract phenotype.30”
From CRISPR-Cas9: a new and promising player in gene therapy (2015) · cites this paper as [30]
In the citing paper
“These results highlighted the importance of HDR in reliable gene corrections.”
From CRISPR-Cas9: a new and promising player in gene therapy (2015) · cites this paper as [30]
In the citing paper
“Of the 22 live offsprings, 10 underwent culprit gene modifications (the editing efficiency is 45.45%) either by HDR (n=4) (figure 1C left) or by NHEJ (n=6) (figure 1C right).”
From CRISPR-Cas9: a new and promising player in gene therapy (2015) · cites this paper as [30]
In the citing paper
“However, as demonstrated in this study, zygote manipulation can only rescue part of the offsprings in an unpredictable manner.”
From CRISPR-Cas9: a new and promising player in gene therapy (2015) · cites this paper as [30]
In the citing paper
“As expected, all of the four offsprings undergoing HDR were free of cataract due to HDR-mediated gene corrections.”
From CRISPR-Cas9: a new and promising player in gene therapy (2015) · cites this paper as [30]
In the citing paper
“As shown in the study by Wu et al 30 among mice genetically edited by CRISPR-Cas9, all HDR-mediated editing corrected the causative gene defects and rescued disease phenotype while only a small part of NHEJ-mediated editing had the corrective effect.”
From CRISPR-Cas9: a new and promising player in gene therapy (2015) · cites this paper as [30]
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“This strategy emerges with a high degree of success to treat the genetic disorders in the CRYGC gene that causes cataracts [14] and the CFTR gene that causes cystic fibrosis [15].”
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BackgroundMethodEngineered Nucleases Lead to Genome Editing Revolution in Rats
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“The correction was noted to occur efficiently with limited off-target modifications, with the resultant mice retaining their fertility and successfully passing on the corrected allele to their offspring [9].”
BackgroundInfluentialShaver, L. G., Sundly, A., & Saif, A. O. (2020) · Journal of Public Health and Primary Care · doi:10.4103/jphpc.jphpc_21_20
ANGPLT3 Impacts Not Only Proteinuria But Also Hyperlipidemia in Nephrotic SyndromePreprint
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“T HE usage of CRISPR-Cas systems for genome editing has been gaining much popularity recently due to the many applications which the technology enables in various domains such as gene therapy and agricultural engineering [1], [2], [3], [4].”
BackgroundCRISPR Cas9 Tip of an Iceberg: It’s Potential in Genome Editing and Many MorePreprint
Ahmad, J., & Siddique, N. (2021) · Preprints.org · doi:10.20944/preprints202107.0182.v1
“The resulted therapy helped in acquiring a cataract-free progeny [29].”
BackgroundGene Editing Through CRISPR-Based Technology
Zhang, Z., Wang, P., & Liu, J. (2022) · Springer eBooks · doi:10.1007/978-981-16-8504-0_2
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Faculty Opinions recommendation of A mouse geneticist's practical guide to CRISPR applications.
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Mohan, S. (2019) · Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature · doi:10.3410/f.736222451.793564125
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Faculty Opinions recommendation of Strategies for Efficient Genome Editing Using CRISPR-Cas9.
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Systemic Overview of Experimental Lens Surgery
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“Since the first time CRISPR/Cas9 system had been used into gene editing(figure 1), it has showed great potential and been widely utilized for anatomizing gene function of disease [38-40] and as a power tool in clinic treatment.”
MethodEmbryons humains génétiquement modifiés : entre progrès scientifique et dilemmes éthiques
Ouzaid, A., Oancea, V.-G., & Ravel, C. (2023) · Éthique & Santé · doi:10.1016/j.etiqe.2023.09.001
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Edycja molekularna a bioetyka chrześcijańska
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Recent Advances in CRISPR-Cas Genome Engineering: An Overview
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Additional file 1 of ANGPTL3 impacts proteinuria and hyperlipidemia in primary nephrotic syndrome
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Repair of leukemia-associated single nucleotide variants via interallelic gene conversionPreprint
Silver, A. J., Brown, D. J., Olmstead, S. D., Watke, J., Gorska, A. E., Tanner, L., Ramsey, H. E., & Savona, M. R. (2024) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2024.04.16.587991
Laoharawee, K., Kleinboehl, E. W., Jensen, J. D., Peterson, J. J., Slipek, N. J., Wick, B., Johnson, M. J., Webber, B. R., & Moriarity, B. (2024) · bioRxiv · doi:10.1101/2024.04.23.590790 · Cited by 2
“21,28,29 Some recent studies have reported results with T cells engineered to produce and secrete various proteins via transposon or lentiviral methods.”
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Progress in developing and implementing CRISPR technologies made by Sun Yat-sen University
Li, Z., Zhou, Y., Huang, J., & Li, J. (2024) · Scientia Sinica Vitae · doi:10.1360/ssv-2024-0145
Gene Modification Technology Using CRISPR/CAS9 System
Okimoto, T., Hino, H., & Shuichi, H. (2024) · Journal of Nihon University Medical Association · doi:10.4264/numa.83.4_121
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Применение технологии CRISPR/Cas9 при наследственных заболеваниях почек
Ван, Ш. (2025) · Рецепт · doi:10.34883/pi.2025.28.2.004
CRISPR-Cas opens a new era of antimicrobial therapy as a powerful gene editing tool
Yang, S., Jiao, X., Liu, J., Liu, Y., Wang, M., Li, S., & Qiao, J. (2025) · World Journal of Microbiology and Biotechnology · doi:10.1007/s11274-025-04591-5
“…of the CRISPR-Cas system (Ishino et al. 1987; Mojica et al. 2005; Grissa et al. 2007; Brouns et al. 2008; Jinek et al. 2012; Cong et al. 2013; Wu et al. 2013 ; Komor et al. 2016; Abu-dayyeh et al. 2016; Gootenberg et al. 2017; Mancuso et al. 2020) RNA from pre-crRNA and trans-activating…”
Synthetic biology and genetic engineering in pharmaceutical bioprospecting
Kant, S., Roy, S., & Sarkar, N. (2025) · Elsevier eBooks · doi:10.1016/b978-0-443-33032-2.00019-8
Metadata
- Authors
- Wu, Y., Liang, D., Wang, Y., Bai, M., Tang, W., Bao, S., Yan, Z., Li, D., & Li, J.
- Affiliations
- Chinese Academy of Sciences (CN)
- Corresponding author
- Li, Jinsong
- Venue
- Cell stem cell
- Publisher
- Elsevier BV
- Published
- 2013-12
- Volume
- 13
- Issue
- 6
- Pages
- 659-662
- ISSN
- 1875-9777 (ISSN-L)
- 1934-5909
- OpenAlex
- W2058792860
- Cited by
- 614
- References
- 20
- Citations by year
- 2026: 14 · 2025: 19 · 2024: 26 · 2023: 34 · 2022: 34 · 2021: 51 · 2020: 51 · 2019: 61 · 2018: 67 · 2017: 64 · 2016: 69 · 2015: 68 · 2014: 54 · 2013: 2
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Cell stem cell (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Animal Genetics and Reproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Keywords
- CRISPR/Cas9, gene correction, cataract, dominant mutations, homology-directed repair, single guide RNA, zygote injection, off-target effects, germline transmission, mouse model
- MeSH terms
- Animals, Base Sequence, Cataract, Disease Models, Animal, Molecular Sequence Data, RNA, Messenger, Genetic Therapy, gamma-Crystallins, Mice, Clustered Regularly Interspaced Short Palindromic Repeats, CRISPR-Associated Proteins
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