Genome editing in cotton with the CRISPR/cas9 system
Open accessAbstract
Genome editing is an important tool for gene functional studies as well as crop improvement. The recent development of the CRISPR/Cas9 system using single guide RNA molecules (sgRNAs) to direct precise double strand breaks in the genome has the potential to revolutionize agriculture. Unfortunately, not all sgRNAs are equally efficient and it is difficult to predict their efficiency by bioinformatics. In crops such as cotton (Gossypium hirsutum L.), with labor-intensive and lengthy transformation procedures, it is essential to minimize the risk of using an ineffective sgRNA that could result in the production of transgenic plants without the desired CRISPR-induced mutations. In this study, we have developed a fast and efficient method to validate the functionality of sgRNAs in cotton using a transient expression system. We have used this method to validate target sites for three different genes GhPDS, GhCLA1, and GhEF1 and analyzed the nature of the CRISPR/Cas9-induced mutations. In our experiments, the most frequent type of mutations observed in cotton cotyledons were deletions (~64%). We prove that the CRISPR/Cas9 system can effectively produce mutations in homeologous cotton genes, an important requisite in this allotetraploid crop. We also show that multiple gene targeting can be achieved in cotton with the simultaneous expression of several sgRNAs and have generated mutations in GhPDS and GhEF1 at two target sites. Additionally, we have used the CRISPR/Cas9 system to produce targeted gene fragment deletions in the GhPDS locus. Finally, we obtained transgenic cotton plants containing CRISPR/Cas9-induced gene editing mutations in the GhCLA1 gene. The mutation efficiency was very high, with 80.6% of the transgenic lines containing mutations in the GhCLA1 target site resulting in an intense albino phenotype due to interference with chloroplast biogenesis.
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Targeting the GhCLA1 gene in cotton improves drought tolerance.
“Furthermore, the application of gene editing techniques extends beyond model plants; for example, targeting the GhCLA1 gene in cotton has been shown to improve drought tolerance (Gao et al., 2017).”
From Emerging applications of gene editing technologies for the development of climate-resilient crops (2025) · cites this paper as Gao et al., 2017
The Results section reports CRISPR/Cas9 editing of GhCLA1 in cotton, supporting the targeting part of the claim. It does not report improved drought tolerance; instead, the edited plantlets showed photobleaching and severely stunted growth. No page number is available in our copy. (LLM, conf=0.88)
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- Authors
- Gao, W., Long, L., Tian, X., Xu, F., Liu, J., Singh, P. K., Botella, J. R., & Song, C.
- Affiliations
- Henan University (CN)
- Cotton Research Institute (CN)
- Venue
- Frontiers in Plant Science
- Publisher
- Frontiers Media SA
- Published
- 2017-08-03
- Volume
- 8
- Pages
- 1364-1364
- ISSN
- 1664-462X (ISSN-L)
- PMC
- PMC5541054
- OpenAlex
- W2744117009
- Cited by
- 254
- References
- 60
- Citations by year
- 2026: 13 · 2025: 27 · 2024: 33 · 2023: 27 · 2022: 28 · 2021: 35 · 2020: 33 · 2019: 30 · 2018: 24 · 2017: 4
- FWCI
- 10.49
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- cc-by
- Locations
- Frontiers in Plant Science (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
- Funders
- National Natural Science Foundation of China (31601344, 31430061)
- Ministry of Agriculture of the People’s Republic of China (2016ZX08009-003)
- 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 Genetic and Mutation Studies
- Keywords
- CRISPR/Cas9 genome editing, cotton, Gossypium hirsutum, transient expression system, homeologous genes, allotetraploid, targeted deletion, transgenic cotton, albino phenotype, chloroplast biogenesis, mutation efficiency
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