Bt Research 2024, Vol.15, No.1, 53-64 http://microbescipublisher.com/index.php/bt 62 microbial strains for the production of biofuels, pharmaceuticals, and other valuable biochemicals (Schulze and Lammers, 2020). The ability to precisely edit metabolic pathways and regulatory networks in industrial microorganisms can lead to more efficient and sustainable bioprocesses. In summary, the future of CRISPR-Cas9 technology in Bt genome editing and functional studies is promising, with emerging trends focusing on increased precision and novel applications in both research and biotechnology. The continued development and refinement of CRISPR tools will undoubtedly drive further innovations and applications in these fields. 9 Concluding Remarks The application of CRISPR-Cas9 technology in Bt genome editing and functional studies has demonstrated significant advancements and potential. The key findings from various studies highlight the efficiency, precision, and versatility of CRISPR-Cas9 in targeted genome editing. This technology has been successfully employed to achieve gene knockouts, regulate endogenous gene expression, and create disease models, thereby expanding the toolkit available for genetic research and therapeutic applications. Despite its transformative impact, challenges such as off-target effects and unintended large gene modifications remain critical concerns. Efforts to mitigate these issues through high-fidelity CRISPR variants and improved sgRNA design are ongoing and show promise in enhancing the specificity and safety of CRISPR-Cas9 applications. The importance of continued research and development in this field cannot be overstated. As we delve deeper into the mechanisms and applications of CRISPR-Cas9, the potential for novel discoveries and technological innovations grows. This ongoing research is crucial for refining the technology, addressing existing limitations, and expanding its applicability to a broader range of organisms and cell types. The development of new Cas effectors and the exploration of microbial diversity are particularly promising areas that could lead to the discovery of new genome editing tools and techniques. Future studies should focus on several key areas to further advance CRISPR-Cas9 technology. First, there is a need for more comprehensive methods to detect and quantify off-target effects and large gene modifications to ensure the safety and efficacy of genome editing. Second, the development of more efficient delivery systems for CRISPR components will be essential for clinical applications. Third, exploring the potential of CRISPR-Cas9 in creating more accurate and diverse disease models will enhance our understanding of genetic disorders and facilitate drug discovery. Finally, interdisciplinary collaborations will be vital in translating CRISPR-Cas9 research from the laboratory to clinical settings, ultimately improving human health and disease treatment. Acknowledgments We wish to express our gratitude to the two anonymous peer reviewers for their valuable feedback on this study's manuscript. Conflict of Interest Disclosure The authors affirm 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.M., 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 Ayanoğlu F.B., Elçin A.E., and Elçin Y.M., 2020, Bioethical issues in genome editing by CRISPR-Cas9 technology, Turkish Journal of Biology, 44(2): 110-120. https://doi.org/10.3906/biy-1912-52 Barman A., Deb B., and Chakraborty S., 2019, A glance at genome editing with CRISPR–Cas9 technology, Current Genetics, 66: 447-462. https://doi.org/10.1007/s00294-019-01040-3 Bishnoi S., 2023, CRISPR-Cas9 gene editing: current progress and future applications, International Journal for Research in Applied Science and Engineering Technology, 21(1): 57. https://doi.org/10.22214/ijraset.2023.55035
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