Cited by 373CRISPR and Genetic EngineeringGenetically Modified Organisms ResearchPlant Molecular Biology Research

CRISPR/Cas: A powerful tool for gene function study and crop improvement

Open access
Zhang, Dangquan, Zhang, Zhiyong, Unver, Turgay, Zhang, Baohong·2021·Journal of Advanced Research

Abstract

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.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1016/j.jare.2020.10.003

Cites

141 of 141 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • AAV delivery in vivo may cause accumulation of off-target mutations.

    “| 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]

    The Future perspectives section (‘Off-target effect is a big concern for CRISPR/Cas application’; page unavailable) says delivery method affects off-target editing and that short-lived RNA or protein delivery reduces the chance of cutting other locations. It does not discuss AAV delivery or say that off-target mutations accumulate during in vivo use. (LLM, conf=0.77)

    Experimental — accuracy not yet measured
  • LNP delivery in vivo eliminates off-target effects.

    “| 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]

    The “Off-target effect is a big concern” section (page not provided) says directly delivered CRISPR/Cas proteins or RNAs, such as RNPs, can be quickly degraded and significantly reduce off-target effects. It does not establish that LNP delivery in vivo eliminates them; the “RNA and protein-level’s delivery” section likewise says RNP transformation may reduce off-target effects. (LLM, conf=0.87)

    Experimental — accuracy not yet measured
  • Designing sgRNA is more convenient, programmable and cost-effective than designing DNA binding domains.

    “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

  • CRISPR/Cas9 is more favored than ZFNs and TALENs and has revolutionized the biotechnology field.

    “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

  • AAV delivery in vivo lasts for years in terminally differentiated cell types.

    “| 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]

    The CRISPR/Cas delivery methods section discusses plant virus vectors, and Future perspectives discusses persistent expression from T-DNA in plants. Neither section addresses AAV delivery or whether it lasts for years in terminally differentiated cells (searched: AAV and adeno-associated virus with those terms). (LLM, conf=0.76)

    Experimental — accuracy not yet measured

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

Flagged first, then by citations

6 of the 147 references couldn’t be matched to a known paper.

Research works printed but not linked: 2

  1. [118]Zhang Y. Efficient and transgene-free genome editing in wheat through transient expression of CRISPR/Cas9 DNA or RNA Nat Commun 7 1 2016 12617 27558837 PMC5007326
  2. [139]Hu J.H. Evolved Cas9 variants with broad PAM compatibility and high DNA specificity Nature 556 7699 2018 57 63 29512652 PMC5951633

Cited by

Sorted by citations

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
OpenAlex
W3094156280
Cited by
373
References
147
Citations by year
2026: 53 · 2025: 95 · 2024: 93 · 2023: 67 · 2022: 46 · 2021: 18 · 2020: 1
FWCI
3.16
Type
journal article
Language
en
Open access
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

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review