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 silico tools are useful to search for potential off-target sites in the whole genome and calculate the likelihood of an off-target editing.
“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
Unwanted mutations may cause a loss of gene function, can be oncogenic or present danger signals that could induce an immune response.
“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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Off-target effects in CRISPR/Cas9 gene editing
Guo, C., Ma, X., Gao, F., & Guo, Y. (2023) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2023.1143157 · Cited by 625 · see citing works
“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).”
BackgroundMechanism and Applications of CRISPR/Cas-9-Mediated Genome Editing
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Comprehensive Overview of Bottom-Up Proteomics Using Mass Spectrometry
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Stimuli-Responsive Gene Delivery Nanocarriers for Cancer Therapy
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Advances in CRISPR therapeutics
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An Outlook on Global Regulatory Landscape for Genome-Edited Crops
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Writers, readers, and erasers RNA modifications and drug resistance in cancer
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Multiplex genome-editing technologies for revolutionizing plant biology and crop improvement
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CRISPR-Based Crop Improvements: A Way Forward to Achieve Zero Hunger
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Current State of Human Gene Therapy: Approved Products and Vectors
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- 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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