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CRISPR/Cas: A powerful tool for gene function study and crop improvement
Open accessAbstract
It is a long-standing goal of scientists and breeders to precisely control a gene for studying its function as well as improving crop yield, quality, and tolerance to various environmental stresses. The discovery and modification of CRISPR/Cas system, a nature-occurred gene editing tool, opens an era for studying gene function and precision crop breeding. In this review, we first introduce the brief history of CRISPR/Cas discovery followed the mechanism and application of CRISPR/Cas system on gene function study and crop improvement. Currently, CRISPR/Cas genome editing has been becoming a mature cutting-edge biotechnological tool for crop improvement that already used in many different traits in crops, including pathogen resistance, abiotic tolerance, plant development and morphology and even secondary metabolism and fiber development. Finally, we point out the major issues associating with CRISPR/Cas system and the future research directions. Key Scientific Concepts of Review: CRISPR/Cas9 system is a robust and powerful biotechnological tool for targeting an individual DNA and RNA sequence in the genome. It can be used to target a sequence for gene knockin, knockout and replacement as well as monitoring and regulating gene expression at the genome and epigenome levels by binding a specific sequence. Agrobacterium-mediated method is still the major and efficient method for delivering CRISPR/Cas regents into targeted plant cells. However, other delivery methods, such as virus-mediated method, have been developed and enhanced the application potentials of CRISPR/Cas9-based crop improvement. PAM requirement offers the CRISPR/Cas9-targted genetic loci and also limits the application of CRISPR/Cas9. Discovering new Cas proteins and modifying current Cas enzymes play an important role in CRISPR/Cas9-based genome editing. Developing a better CRISPR/Cas9 system, including the delivery system and the methods eliminating off-target effects, and finding key/master genes for controlling crop growth and development is two major directions for CRISPR/Cas9-based crop improvement.
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![Fig. 2 Application of CRISPR/Cas genome editing in crop improvement, CRISPR/Cas is a robust tool and have huge potentials on crop improvement. Though targeting an individual DNA sequence, CRISPR/Cas can be used to add and/or change the expression of an individual gene that controls plant tolerance to abiotic and biotic stresses as well as plant growth and development. CRISPR/Cas also can change the secondary metabolism for controlling the biosynthesis of protein, carbohydrate, oil, and functional components. Partial figures are modified from previous publications [74], [108].](/paper-figures/6f08ea3b35d948ec97e53b01adce3ae0/crispr-cas-a-powerful-tool-for-gene-function-study-and-crop-improvement-figure-2.jpg)
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“| Delivery methods improvement | Application in vivo [95–105] | AAV: Last for years in terminally differentiated cell types | May cause accumulation of off-target mutations |”
From Off-target effects in CRISPR/Cas9 gene editing (2023) · cites this paper as [101]
Experimental — accuracy not yet measuredThe source backs part of the claim, not all of it — or backs it for a different population, condition or comparison.
In the citing paper
“| Delivery methods improvement | Application in vivo [95–105] | LNP: Can be quickly degraded in vivo | Eliminate off-target effects |”
From Off-target effects in CRISPR/Cas9 gene editing (2023) · cites this paper as [101]
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“Designing sgRNA is more convenient, programmable and cost-effective than designing DNA binding domains, thus CRISPR/Cas9 is more favored than ZFNs and TALENs and has revolutionized the biotechnology field (Zhang et al., 2021; Tran et al., 2022).”
From Off-target effects in CRISPR/Cas9 gene editing (2023) · cites this paper as Zhang et al., 2021
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“Designing sgRNA is more convenient, programmable and cost-effective than designing DNA binding domains, thus CRISPR/Cas9 is more favored than ZFNs and TALENs and has revolutionized the biotechnology field (Zhang et al., 2021; Tran et al., 2022).”
From Off-target effects in CRISPR/Cas9 gene editing (2023) · cites this paper as Zhang et al., 2021
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In the citing paper
“| Delivery methods improvement | Application in vivo [95–105] | AAV: Last for years in terminally differentiated cell types | May cause accumulation of off-target mutations |”
From Off-target effects in CRISPR/Cas9 gene editing (2023) · cites this paper as [101]
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In the citing paper
“| Delivery methods improvement | Application in vivo [95–105] | LNP: Can be quickly degraded in vivo | Eliminate off-target effects |”
From Off-target effects in CRISPR/Cas9 gene editing (2023) · cites this paper as [101]
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Metadata
- Authors
- Zhang, D., Zhang, Z., Unver, T., & Zhang, B.
- Affiliations
- Henan Agricultural University (CN)
- Henan Institute of Science and Technology (CN)
- East Carolina University (US)
- Corresponding author
- Zhang, Dangquan
- Zhang, Zhiyong
- Zhang, Baohong
- Venue
- Journal of Advanced Research
- Publisher
- Elsevier BV
- Published
- 2021-03
- Volume
- 29
- Pages
- 207-221
- ISSN
- 2090-1224 (ISSN-L)
- 2090-1232
- PMC
- PMC8020163
- OpenAlex
- W3094156280
- Cited by
- 373
- References
- 147
- Citations by year
- 2026: 53 · 2025: 95 · 2024: 93 · 2023: 67 · 2022: 46 · 2021: 18
- FWCI
- 3.16
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Journal of Advanced Research (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- National Natural Science Foundation of China (31172257, 31571600, 31271648)
- Science and Technology Innovation Talents in Universities of Henan Province (114100510008)
- Cotton Incorporated
- National Science Foundation (1658709)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Agricultural and Biological Sciences › Plant Science › Genetically Modified Organisms Research
- Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant Molecular Biology Research
- Keywords
- CRISPR/Cas9, genome editing, gene function, crop improvement, precision breeding, pathogen resistance, abiotic stress tolerance, plant development, secondary metabolism, fiber development, protospacer adjacent motif, off-target effects, Cas proteins, gene knock-in, gene knockout, epigenome editing
- MeSH terms
- Plant Breeding, Gene Editing, Agriculture, Humans, Phenotype, Stress, Physiological, Gene Targeting, Crops, Agricultural, Genome, Plant, Plants, Genetically Modified, Gene Knock-In Techniques, Agrobacterium, Plant Development, Clustered Regularly Interspaced Short Palindromic Repeats, CRISPR-Cas Systems
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