Phage and CRISPR based precision antimicrobials: a dual strategy against multidrug-resistant bacteria

Artificial intelligence and CRISPR-based approaches for targeted delivery of bacteriophages.

The rapid emergence of multidrug-resistant (MDR) bacteria has increased interest in bacteriophage therapy as a promising alternative to conventional antibiotics. Bacteriophages are host-specific bacterial viruses that selectively infect and destroy pathogenic bacterial strains. Recent developments in artificial intelligence (AI) and CRISPR-based technologies offer innovative approaches to address challenges such as narrow host range, rapid immune clearance, phage instability, bacterial resistance, and biofilm penetration barriers. By integrating AI-driven structural modeling with CRISPR-mediated genome editing, these methods enable the targeted delivery of bacteriophages. This review focuses on next-generation approaches that combine AI-assisted phage identification, host prediction, and therapeutic optimization with CRISPR-based genome engineering for targeted phage delivery and improved safety. Overall, this review highlights the potential of AI- and CRISPR-assisted phage therapy for the treatment of MDR bacterial infections. This review provides a systematic overview of bacteriophage biology, life cycle, and mechanisms of action, highlighting the influence of phage morphology on therapeutic performance, recent advances, current clinical and preclinical studies, and future perspectives. Although phage therapy shows considerable potential against MDR bacterial infections, several challenges related to delivery, safety, and clinical translation remain. The integration of AI and CRISPR technologies may improve phage selection, targeting specificity, and therapeutic performance. Continued research, clinical validation, and regulatory development will be essential for translating these advances into practical antimicrobial therapies.

CRISPR/Cas system as a novel therapeutic strategy to combat multi-drug-resistant bacteria

Phage Therapy Enhanced by Using Engineered Bacteriophages: A Powerful Antibacterial Tool to Address the Dilemma Posed by Multidrug-Resistant Bacterial Infections

The continuous slowdown in the research and development of new antibiotics and antibiotic overuse have turned the problem of antibacterial resistance into a global public health crisis. As a very promising alternative to multi-drug-resistant bacterial infection, phage therapy is receiving renewed attention. However, the inherent biological limitations of natural phages restrict their extensive clinical application. This review examines how synthetic biology can be harnessed to transform phages and to build the next generation of antibacterial therapies. We outline the main advantages of natural phages, including high host specificity, self-amplification, bactericidal activity and the ability to degrade biofilms. We also point out the bottlenecks of clinical applications of bacteriophages, such as narrow host range, rapid removal in the body and potential genetic safety risks. Moreover, we elaborate on the core synthetic biological tools used to overcome the above limitations, including CRISPR-Cas gene editing, receptor-binding protein reprogramming, functional load delivery and immunogenic regulation, and summarize the recent clinical progress and personalized treatment process. The increasing clinical evidence shows that synthetic biology can effectively overcome the inherent defects of natural bacteriophages, confirming the safety and initial efficacy of bacteriophage therapy. Engineered phages provide a practical strategy to meet the antimicrobial resistance challenge. Clinical applications of such phages will mainly depend on progress in production standardization, regulatory framework construction and scientific and reasonable joint treatment program development.

CRISPR-ENABLED ANTIMICROBIAL STRATEGIES: REDEFINING PRECISION THERAPY AGAINST MULTIDRUG RESISTANT PATHOGENS

Antimicrobial resistance (AMR) has become one of the most serious global public health threats, driven by the rapid emergence and dissemination of multidrug-resistant (MDR) pathogens. The declining effectiveness of conventional antibiotics necessitates the development of innovative therapeutic strategies capable of selectively targeting resistant microorganisms while preserving beneficial microbiota. CRISPR-Cas technology has emerged as a promising precision antimicrobial platform with the potential to transform infectious disease management. This review summarizes the molecular mechanisms, delivery platforms, therapeutic applications, emerging innovations, safety considerations, translational barriers, and future prospects of CRISPR-enabled antimicrobial strategies for combating MDR bacterial infections. This review evaluated recent peer-reviewed literature on CRISPR-Cas systems, AMR, genome editing, precision medicine, delivery technologies, and translational research. The available evidence was synthesized to assess current advances, therapeutic potential, existing limitations, and future opportunities for CRISPR-based antimicrobial interventions. CRISPR-enabled antimicrobial strategies demonstrate remarkable specificity by selectively eliminating multidrug-resistant bacteria, disrupting resistance genes, targeting virulence factors, inhibiting biofilm formation, and restoring antibiotic susceptibility. Advances in bacteriophage-mediated delivery, nanoparticle-based carriers, programmable RNA-targeting systems, artificial intelligence-assisted guide RNA design, synthetic biology, and personalized precision medicine have further expanded their clinical potential. Nevertheless, challenges related to delivery efficiency, off-target editing, bacterial escape mechanisms, biosafety, regulatory approval, and large-scale manufacturing remain significant translational barriers. Overall, CRISPR-based antimicrobial therapy represents a transformative approach to precision infectious disease management, with continued technological innovation, interdisciplinary collaboration, and robust clinical validation expected to facilitate its safe, effective, and clinically accessible implementation against multidrug-resistant pathogens.

Bacteriophage Therapy as a Promising Substitute for Antibiotics: Current Advances, Clinical Applications, and Future Perspectives

The rapid emergence and global dissemination of antimicrobial-resistant (AMR) bacteria have significantly reduced the effectiveness of conventional antibiotics, posing a major threat to modern healthcare. The increasing prevalence of multidrug-resistant pathogens, coupled with the slow development of new antimicrobial agents, has renewed interest in alternative therapeutic strategies. Among these, bacteriophage (phage) therapy has re-emerged as a promising biological approach due to its ability to selectively infect and eliminate bacterial pathogens while preserving the normal microbiota. Unlike broad- spectrum antibiotics, bacteriophages exhibit high host specificity, self-replicate at the site of infection, and can effectively disrupt bacterial biofilms, making them particularly attractive for treating chronic and drug-resistant infections. This review critically examines the biological characteristics of bacteriophages, their mechanisms of antibacterial action, therapeutic applications, advantages, and current limitations. Recent advances in phage engineering, genome editing, phage cocktails, encapsulation technologies, and combination therapies with antibiotics are also discussed, highlighting their potential to improve treatment efficacy and overcome bacterial resistance. Furthermore, the review evaluates findings from recent preclinical and clinical studies, regulatory challenges, manufacturing considerations, and safety issues that currently limit the widespread clinical implementation of phage therapy. Although several scientific and regulatory hurdles remain, accumulating experimental and clinical evidence indicates that bacteriophage therapy could become an important component of future antimicrobial strategies. Continued multidisciplinary research, standardized clinical protocols, and well-designed randomized clinical trials are essential to establish its long-term efficacy and safety. With ongoing advances in molecular biology, synthetic biology, and precision medicine, bacteriophage therapy has the potential to complement or, in selected clinical situations, replace conventional antibiotics in the management of multidrug-resistant bacterial infections.

Plasmid-mediated multidrug resistance and the antibacterial potential of plant seed proteins

The rapid emergence of multidrug-resistant (MDR) bacteria, largely driven by excessive and indiscriminate antibiotic use, poses a critical threat to global health. Conventional antibiotics are increasingly ineffective against resistant pathogens, particularly those harboring plasmid-mediated resistance mechanisms. The purpose of this study is to assess the antibacterial efficacy of chickpea-derived seed proteins (7S, 11S globulins, and bioactive seed fractions) as natural alternative treatments, and to determine the prevalence of MDR among clinical bacterial isolates. We hypothesized that chickpea seed proteins, particularly the basic subunit (BS) fraction, would exhibit significant antibacterial efficacy against MDR strains, including those harboring plasmid-mediated resistance. Ten clinical bacterial isolates were subjected to antibiotic susceptibility testing using the Kirby–Bauer disc diffusion method. Resistant strains were further analyzed through plasmid profiling, curing experiments, and agarose gel electrophoresis to distinguish plasmid-mediated from chromosomal resistance. Antibacterial proteins were isolated from chickpea seeds, including 7S and 11S globulins, along with a BS fraction. Their antibacterial activity was assessed against resistant isolates. Using scanning electron microscopy (SEM) and transmission electron microscopy (TEM), morphological and structural alterations brought on by protein treatment were investigated, offering insights into bacterial adaptability and protein-mediated inhibition. Antibiotic susceptibility testing revealed widespread resistance, with cephalexin and clindamycin showing the lowest effectiveness (80–90% resistance). Klebsiella oxytoca and Proteus mirabilis exhibited complete resistance (Multi antibiotics resistant [MAR] = 1.0), underscoring their clinical significance. Plasmid profiling demonstrated that 13 of 21 resistant isolates carried plasmids, confirming plasmid-mediated resistance to β-lactam and phenicol antibiotics, while others exhibited chromosomal resistance. TEM and SEM analyses revealed that resistant Klebsiella oxytoca and Bacillus subtilis maintained intact morphology despite high antibiotic exposure, suggesting mutation-driven adaptation. Chickpea-derived protein fractions displayed notable antibacterial activity, with the BS fraction exerting the strongest inhibition, particularly against K. oxytoca and P. mirabilis . This study highlights the alarming prevalence of plasmid- and mutation-mediated antibiotic resistance among clinical isolates. Chickpea proteins, especially the BS fraction, demonstrated significant antibacterial activity against MDR pathogens, offering a promising natural alternative to conventional antibiotics. Their ability to inhibit resistant strains suggests potential applications in therapeutic development and infection control.