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Latest Developed Strategies to Minimize the Off-Target Effects in CRISPR-Cas-Mediated Genome Editing
Open accessAbstract
Gene editing that makes target gene modification in the genome by deletion or addition has revolutionized the era of biomedicine. Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 emerged as a substantial tool due to its simplicity in use, less cost and extraordinary efficiency than the conventional gene-editing tools, including zinc finger nucleases (ZFNs) and Transcription activator-like effector nucleases (TALENs). However, potential off-target activities are crucial shortcomings in the CRISPR system. Numerous types of approaches have been developed to reduce off-target effects. Here, we review several latest approaches to reduce the off-target effects, including biased or unbiased off-target detection, cytosine or adenine base editors, prime editing, dCas9, Cas9 paired nickase, ribonucleoprotein (RNP) delivery and truncated gRNAs. This review article provides extensive information to cautiously interpret off-target effects to assist the basic and clinical applications in biomedicine.
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“In this scenario, in silico tools are useful to search for potential off-target sites in the whole genome and calculate the likelihood of an off-target editing (Naeem et al., 2020).”
From Off-target effects in CRISPR/Cas9 gene editing (2023) · cites this paper as Naeem et al., 2020
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“Generally speaking, studies revealed that GC content of between 40% and 60%, truncated (short length of sgRNA), and incorporation of 2ʹ-O-methyl-3ʹ-phosphonoacetate in the sgRNA ribose-phosphate backbone are the preferred methods to increase genome editing efficiency of CRISPR/Cas-9.67,68”
From Mechanism and Applications of CRISPR/Cas-9-Mediated Genome Editing (2021) · cites this paper as [68]
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“Generally speaking, studies revealed that GC content of between 40% and 60%, truncated (short length of sgRNA), and incorporation of 2ʹ-O-methyl-3ʹ-phosphonoacetate in the sgRNA ribose-phosphate backbone are the preferred methods to increase genome editing efficiency of CRISPR/Cas-9.67,68”
From Mechanism and Applications of CRISPR/Cas-9-Mediated Genome Editing (2021) · cites this paper as [68]
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“These unwanted mutations may cause a loss of gene function, can be oncogenic or present danger signals that could induce an immune response [109].”
From Immune responses to gene editing by viral and non-viral delivery vectors used in retinal gene therapy (2022) · cites this paper as [109]
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“Generally speaking, studies revealed that GC content of between 40% and 60%, truncated (short length of sgRNA), and incorporation of 2ʹ-O-methyl-3ʹ-phosphonoacetate in the sgRNA ribose-phosphate backbone are the preferred methods to increase genome editing efficiency of CRISPR/Cas-9.67,68”
From Mechanism and Applications of CRISPR/Cas-9-Mediated Genome Editing (2021) · cites this paper as [68]
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Metadata
- Authors
- Naeem, M., Majeed, S., Hoque, M. Z., & Ahmad, I.
- Affiliations
- King Fahd University of Petroleum and Minerals (SA)
- Texas Tech University (US)
- Corresponding author
- Ahmad, Irshad
- Venue
- Cells
- Publisher
- MDPI AG
- Published
- 2020-07-02
- Volume
- 9
- Issue
- 7
- Pages
- 1608
- ISSN
- 2073-4409 (ISSN-L)
- PMC
- PMC7407193
- OpenAlex
- W3038956601
- Cited by
- 457
- References
- 156
- Citations by year
- 2026: 33 · 2025: 73 · 2024: 93 · 2023: 89 · 2022: 89 · 2021: 73 · 2020: 7
- FWCI
- 6.17
- Type
- journal article
- Language
- en
- Open access
- gold
- dx.doi.org
- Licence
- Locations
- Cells (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- King Fahd University of Petroleum and Minerals (31261)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Keywords
- CRISPR/Cas9, off-target effects, genome editing, off-target detection, base editors, prime editing, dCas9, Cas9 nickase, ribonucleoprotein delivery, zinc finger nucleases, TALEN
- MeSH terms
- Gene Editing, CRISPR-Associated Protein 9, RNA, Guide, CRISPR-Cas Systems, Animals, DNA Repair, Humans, Ribonucleoproteins, RNA, Guide, Kinetoplastida, CRISPR-Cas Systems
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