Legume Genomics and Genetics 2024, Vol.15, No.4, 199-209 http://cropscipublisher.com/index.php/lgg 207 delivery methods and high-throughput mutant libraries are expected to enhance the efficiency and applicability of CRISPR/Cas9 in legume breeding. While the potential of CRISPR/Cas9 in legume breeding is immense, several challenges need to be addressed to fully realize its benefits. Regulatory hurdles and public acceptance remain significant barriers, particularly in regions with stringent regulations on genetically modified organisms. Additionally, the efficiency of transformation and regeneration in certain legume species needs to be improved to make the technology more broadly applicable. Despite these challenges, the ongoing advancements in CRISPR/Cas9 technology, including the development of more efficient delivery systems and the refinement of gene-editing techniques, offer promising solutions. The continued research and collaboration among scientists, regulatory bodies, and the public will be crucial in harnessing the full potential of CRISPR/Cas9 for the sustainable improvement of legume crops. Acknowledgments The author is grateful to the two anonymous peer reviewers for their insightful feedback on the manuscript. Conflict of Interest Disclosure The author affirms that this research was conducted without any commercial or financial relationships that could be construed as a potential conflict of interest. References Abdelrahman M., Al-Sadi A., Pour-Aboughadareh A., Burritt D., and Tran L., 2018, Genome editing using CRISPR/Cas9-targeted mutagenesis: an opportunity for yield improvements of crop plants grown under environmental stresses, Plant Physiology and Biochemistry, 131: 31-36. https://doi.org/10.1016/j.plaphy.2018.03.012 Arora L., and Narula A., 2017, Gene editing and crop improvement using CRISPR-Cas9 system, Frontiers in Plant Science, 8: 1932. https://doi.org/10.3389/fpls.2017.01932 Badhan S., Ball A., and Mantri N., 2021, First report of CRISPR/Cas9 mediated DNA-free editing of 4CL and RVE7 genes in chickpea protoplasts, International Journal of Molecular Sciences, 22(1): 396. https://doi.org/10.3390/ijms22010396 Baloğlu M., Altunoğlu Y., Baloglu P., Yildiz A., Türkölmez N., and Çiftçi Y., 2022, Gene-editing technologies and applications in legumes: progress, evolution, and future prospects, Frontiers in Genetics, 13: 859437. https://doi.org/10.3389/fgene.2022.859437 Belhaj K., Chaparro‐Garcia A., Kamoun S., Patron N., and Nekrasov V., 2015, Editing plant genomes with CRISPR/Cas9, Current Opinion in Biotechnology, 32: 76-84. https://doi.org/10.1016/j.copbio.2014.11.007 Bhowmik P., Konkin D., Polowick P., Hodgins C., Subedi M., Xiang D., Yu B., Patterson N., Rajagopalan N., Babic V., Ro D., Tar’an B., Bandara M., Smyth S., Cui Y., and Kagale S., 2021, CRISPR/Cas9 gene editing in legume crops: opportunities and challenges, Legume Science, 3(3): e96. https://doi.org/10.1002/leg3.96 Cai Y., Chen L., Zhang Y., Yuan S., Su Q., Sun S., Wu C., Yao W., Han T., and Hou W., 2020, Target base editing in soybean using a modified CRISPR/Cas9 system, Plant Biotechnology Journal, 18: 1996-1998. https://doi.org/10.1111/pbi.13386 Chen K., Wang Y., Zhang R., Zhang H., and Gao C., 2019, CRISPR/Cas genome editing and precision plant breeding in agriculture, Annual Review of Plant Biology, 70: 667-697. https://doi.org/10.1146/annurev-arplant-050718-100049 Concordet J., and Haeussler M., 2018, CRISPOR: intuitive guide selection for CRISPR/Cas9 genome editing experiments and screens, Nucleic Acids Research, 46: W242-W245. https://doi.org/10.1093/nar/gky354 Corte L., Mahmoud L., Moraes T., Mou Z., Grosser J., and Dutt M., 2019, Development of improved fruit, vegetable, and ornamental crops using the CRISPR/Cas9 genome editing technique, Plants, 8(12): 601. https://doi.org/10.3390/plants8120601 Do P., Nguyen C., Bui H., Tran L., Stacey G., Gillman J., Zhang Z., and Stacey M., 2019, Demonstration of highly efficient dual gRNA CRISPR/Cas9 editing of the homeologous GmFAD2-1Aand GmFAD2-1Bgenes to yield a high oleic, low linoleic and α-linolenic acid phenotype in soybean, BMC Plant Biology, 19: 1-14. https://doi.org/10.1186/s12870-019-1906-8 Fu Y., Sander J., Reyon D., Cascio V., and Joung J., 2014, Improving CRISPR-Cas nuclease specificity using truncated guide RNAs, Nature Biotechnology, 32: 279-284. https://doi.org/10.1038/nbt.2808
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