Bioscience Methods 2024, Vol.15, No.5, 216-225 http://bioscipublisher.com/index.php/bm 223 provided in vivo evidence of the roles of ABCC2 and ABCC3 proteins as functional receptors for Bt Cry1 toxins, offering insights into the molecular mechanisms of insect resistance. The integration of Bt genes into other bacterial strains, such as Bacillus suBtilis, has also been explored, resulting in enhanced biocontrol efficacy against both insect pests and plant pathogens. Furthermore, whole genome sequencing of various Bt strains has revealed a plethora of pesticidal protein genes, underscoring the genetic diversity and potential for biotechnological applications. CRISPR technology has revolutionized the field of Bt insecticides by enabling precise genetic modifications that enhance the efficacy and specificity of Bt proteins. The ability to knockout specific genes, such as PxABCC2 and PxABCC3, has provided a deeper understanding of the genetic basis of resistance in target pests, facilitating the development of more effective Bt strains. This technology has also allowed for the creation of novel chimeric proteins with improved insecticidal properties, as seen with Cry1A.2 and Cry1B.2, which offer new tools for pest management. The integration of CRISPR with Bt technology not only enhances the insecticidal spectrum but also addresses the issue of resistance, thereby ensuring the long-term sustainability of Bt-based biopesticides. The advancements in CRISPR-based gene editing of Bt have the potential to significantly impact global agricultural practices. By developing Bt strains with enhanced insecticidal properties and reduced resistance, farmers can achieve more effective pest control, leading to higher crop yields and reduced reliance on chemical pesticides. This is particularly important for major crops such as soybean and maize, where pest resistance has been a significant challenge. The use of genetically engineered Bt strains can also contribute to more sustainable agricultural practices by reducing the environmental impact of pest control measures and promoting the use of eco-friendly biopesticides. Overall, the integration of CRISPR technology with Bt insecticides holds great promise for improving pest management strategies and supporting global food security. Acknowledgments The author thanks the anonymous peer review for their critical comments and revising suggestion. 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. Reference Ahmad N., Rahman M., Mukhtar Z., Zafar Y., and Zhang B., 2020, A critical look on CRISPR‐based genome editing in plants, Journal of Cellular Physiology, 235(2): 666-682. https://doi.org/10.1002/jcp.29052 Arsov A., Gerginova M., Paunova-Krasteva T., Petrov K., and Petrova P., 2023, Multiple cry genes in Bacillus thuringiensis strain BTG suggest a broad-spectrum insecticidal activity, International Journal of Molecular Sciences, 24(13): 11137. https://doi.org/10.3390/ijms241311137 Bao A., Burritt D., Chen H., Zhou X., Cao D., and Tran L., 2019, The CRISPR/Cas9 system and its applications in crop genome editing, Critical Reviews in Biotechnology, 39(3): 321-336. https://doi.org/10.1080/07388551.2018.1554621 Bisht D., Bhatia V., and Bhattacharya R., 2019, Improving plant-resistance to insect-pests and pathogens: the new opportunities through targeted genome editing, Seminars in Cell & Developmental Biology, 96: 65-76. https://doi.org/10.1016/j.semcdb.2019.04.008 Cao B., Shu C., Geng L., Song F., and Zhang J., 2020, Cry78Ba1, One novel crystal protein fromBacillus thuringiensis with high insecticidal activity against rice planthopper, Journal of Agricultural and Food Chemistry, 68(8): 2539-2546. https://doi.org/10.1021/acs.jafc.9b07429 Chen D., Moar W., Jerga A., Gowda A., Milligan J., Bretsynder E., Rydel T., Baum J., Semeão A., Fu X., Guzov V., Gabbert K., Head G., and Haas J., 2021, Bacillus thuringiensis chimeric proteins Cry1A.2 and Cry1B.2 to control soybean lepidopteran pests: new domain combinations enhance insecticidal spectrum of activity and novel receptor contributions, PLoS ONE, 16(6): e0249150. https://doi.org/10.1371/journal.pone.0249150 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(1): 667-697. https://doi.org/10.1146/annurev-arplant-050718-100049
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