Abstract:The widespread use of antibiotics has exacerbated the multidrug resistance of bacteria, posing a grave threat to public health. Gene knockout, as a precise method for intervening in bacterial genetics, exhibits immense potential in deciphering the mechanisms of multidrug resistance and exploring novel therapeutic approaches. This paper reviews the advancements in the application of gene knockout in the research on multidrug-resistant bacteria. We discovered that gene knockout not only facilitates the understanding of the drug resistance mechanisms in multidrug-resistant bacteria, but also provides crucial insights into identifying new drug targets. We further discuss the advantages and limitations of this technology and propose potential solutions. The further optimization of gene knockout and the integration of this technology with other biotechnologies will provide more effective strategies for addressing the multidrug-resistance in bacteria. We hope that gene knockout can be applied in a broader scope in clinical practice, offering new possibilities for the treatment of multidrug-resistant bacteria.