CRISPR/Cas9 Mechanism and Applications Progression
DOI:
https://doi.org/10.62051/b7416e56Keywords:
CRISPR; Cas9; gene knock-out; Alzheimer’s; Sickle cell disease.Abstract
CRISPR (short for Clustered Regularly Interspaced Short Palindromic Repeats) and the accompanying endonuclease Cas9 are potent tools for gene editing in many applied disciplines. The CRISPR/Cas9 system primarily functions through two components, in which the single guide RNA (sgRNA) directs the Cas9 protein with endonuclease activity to carry out precise DNA cleavage. The cell's inherent repair mechanisms allow it to complete the subsequent reconnection of the broken genes. Gene knock-in or knock-out can therefore be achieved via this approach. This system was typically applied to prokaryotes in early studies. Its role as a component of their defense mechanism has been thoroughly studied. With rapid development over the past ten years, this system can now be effectively used in a multitude of eukaryotic cells, including yeast cells and mammalian cells. Because of its accuracy, effectiveness, and low cost, CRISPR/Cas9 has been widely used for human disease treatment and has shown great potential in gene therapies for hereditary diseases. However, since more late-stage clinical trials are required to validate its safety, its applications still face many challenges. This review investigates the mechanism of CRISPR/Cas9-mediated gene editing technology and highlights its recent applications in Alzheimer's disease, Sickle cell disease, and Duchenne muscular dystrophy, which are difficult to treat with current approaches. Some challenges the CRISPR/Cas9 system has encountered at this stage are described, and its prospects are also examined.
Downloads
References
Li, T., Yang, Y., Qi, H., Cui, W., Zhang, L., Fu, X., He, X., Liu, M., Li, P., et al., “CRISPR/Cas9 Therapeutics: Progress and Prospects,” Signal Transduction and Targeted Therapy, 8 (1), (2023).
Mengstie, M.A., and Wondimu, B.Z., “Mechanism and applications of CRISPR/cas-9-mediated genome editing,” Biologics: Targets and Therapy, Volume 15, 353 – 361 (2021).
Zhang, Y.., Yin, H., “Genome editing with mrna encoding ZFN, Talen, and cas9,” Molecular Therapy, 27 (4), 735 – 746 (2019).
Xu, X., Liu, C., Wang, Y., Koivisto, O., Zhou, J., Shu, Y., and Zhang, H., “Nanotechnology-based delivery of CRISPR/Cas9 for cancer treatment,” Advanced Drug Delivery Reviews, 176, 113891 (2021).
Feng, Y., Liu, S., Chen, R., and Xie, A., “Target binding and residence: A new determinant of DNA double-strand break repair pathway choice in CRISPR/cas9 genome editing,” Journal of Zhejiang University-SCIENCE B, 22 (1), 73 – 86 (2021).
Lu, L., Yu, X., Cai, Y., Sun, M., and Yang, H., “Application of CRISPR/Cas9 in alzheimer’s disease,” Frontiers in Neuroscience, 15, (2021).
Bhardwaj, S., Kesari, K.K., Rachamalla, M., Mani, S., Ashraf, G.Md., Jha, S.K., Kumar, P., Ambasta, R.K., Dureja, H., et al., “CRISPR/Cas9 gene editing: New hope for alzheimer’s disease therapeutics,” Journal of Advanced Research, 40, 207 – 221 (2022).
Sun, J., Carlson-Stevermer, J., Das, U., Shen, M., Delenclos, M., Snead, A.M., Koo, S.Y., Wang, L., Qiao, D., et al., “CRISPR/Cas9 editing of App C-terminus attenuates β-cleavage and promotes α-cleavage,” Nature Communications, 10 (1), (2019).
Wadhwani, A.R., Affaneh, A., Van Gulden, S., and Kessler, J.A., “Neuronal apolipoprotein E4 increases cell death and phosphorylated tau release in alzheimer disease,” Annals of Neurology, 85 (5), 726 – 739 (2019).
Park, S.H., and Bao, G., “CRISPR/Cas9 gene editing for curing sickle cell disease,” Transfusion and Apheresis Science, 60 (1), 103060 (2021).
Demirci, S., Leonard, A., Essawi, K., and Tisdale, J.F., “CRISPR-Cas9 to induce fetal hemoglobin for the treatment of sickle cell disease,” Molecular Therapy - Methods & Clinical Development, 23, 276 – 285 (2021).
Wu, Y., Zeng, J., Roscoe, B.P., Liu, P., Yao, Q., Lazzarotto, C.R., Clement, K., Cole, M.A., Luk, K., et al., “Highly efficient therapeutic gene editing of human hematopoietic stem cells,” Nature Medicine, 25 (5), 776 – 783 (2019).
Wang, L., Li, L., Ma, Y., Hu, H., Li, Q., Yang, Y., Liu, W., Yin, S., Li, W., et al., “Reactivation of γ-globin expression through cas9 or base editor to treat β-hemoglobinopathies,” Cell Research, 30 (3), 276 – 278 (2020).
Happi Mbakam, C., Lamothe, G., Tremblay, G., and Tremblay, J.P., “CRISPR-Cas9 gene therapy for Duchenne muscular dystrophy,” Neurotherapeutics, 19 (3), 931 – 941 (2022).
Mollanoori, H., Rahmati, Y., Hassani, B., Havasi Mehr, M., and Teimourian, S., “Promising therapeutic approaches using CRISPR/cas9 genome editing technology in the treatment of Duchenne muscular dystrophy,” Genes & Diseases, 8 (2), 146 – 156 (2021).
Min, Y.-L., Li, H., Rodriguez-Caycedo, C., Mireault, A.A., Huang, J., Shelton, J.M., McAnally, J.R., Amoasii, L., Mammen, P.P., et al., “CRISPR-Cas9 corrects duchenne muscular dystrophy exon 44 deletion mutations in mice and human cells,” Science Advances, 5 (3), (2019).
Amoasii, L., Hildyard, J.C., Li, H., Sanchez-Ortiz, E., Mireault, A., Caballero, D., Harron, R., Stathopoulou, T.-R., Massey, C., et al., “Gene editing restores dystrophin expression in a canine model of Duchenne muscular dystrophy,” Science, 362 (6410), 86 – 91 (2018).
Downloads
Published
Conference Proceedings Volume
Section
License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
