Microbial biotechnology and genetic engineering play crucial roles in developing therapeutic agents, including antibiotics, anticancer drugs, vaccines, and therapeutic enzymes. Genetic engineering techniques, functional genomics, and synthetic biology have unlocked previously unknown natural products. This review highlights significant advances in microbial biotechnology, focusing on gene-based technologies for medical applications. Microbial products are essential for producing various therapeutic agents, such as antibiotics, antifungals, antivirals, and antiparasitic agents. Genetic engineering has revolutionized the field by enabling the creation of new and diverse therapeutic compounds. The discovery of CRISPR-Cas components has transformed genome editing, enhancing the biotechnological capabilities of specific microorganisms. In medicine, these technologies extend beyond therapeutic compound development to impact diagnosis, prevention, gene expression regulation, and the construction of medical devices. The review also discusses the historical context of drug discovery, the classification and mechanisms of action of antibiotics, and the use of genetic engineering to enhance antibiotic production and create new diversity. Additionally, it covers other bioactive compounds produced by bacteria, such as antifungals, antivirals, and antiparasitic agents, and the challenges and opportunities in their development.Microbial biotechnology and genetic engineering play crucial roles in developing therapeutic agents, including antibiotics, anticancer drugs, vaccines, and therapeutic enzymes. Genetic engineering techniques, functional genomics, and synthetic biology have unlocked previously unknown natural products. This review highlights significant advances in microbial biotechnology, focusing on gene-based technologies for medical applications. Microbial products are essential for producing various therapeutic agents, such as antibiotics, antifungals, antivirals, and antiparasitic agents. Genetic engineering has revolutionized the field by enabling the creation of new and diverse therapeutic compounds. The discovery of CRISPR-Cas components has transformed genome editing, enhancing the biotechnological capabilities of specific microorganisms. In medicine, these technologies extend beyond therapeutic compound development to impact diagnosis, prevention, gene expression regulation, and the construction of medical devices. The review also discusses the historical context of drug discovery, the classification and mechanisms of action of antibiotics, and the use of genetic engineering to enhance antibiotic production and create new diversity. Additionally, it covers other bioactive compounds produced by bacteria, such as antifungals, antivirals, and antiparasitic agents, and the challenges and opportunities in their development.