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

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

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

This review critically examines bacteriophage-based antimicrobials, CRISPR-Cas therapeutic systems, and their emerging integration as CRISPR-armed phages, highlighting their comparative advantages, current limitations, and future potential as promising targeted antimicrobial approach platforms requiring further clinical validation.

Antibacterial Immunotherapy: Mechanistic Insights, Emerging Therapeutic Strategies, and Clinical Translation

Abstract Antimicrobial resistance (AMR) continues to compromise the effectiveness of conventional antibacterial therapy, driving the development of therapeutic strategies that extend beyond direct antibiotic-mediated bacterial killing. Multidrug-resistant (MDR) pathogens evade treatment through diverse mechanisms, including enzymatic drug inactivation, target modification, efflux pump overexpression, biofilm formation, and persisters development. AMR results in chronic and recurrent infections, prolonged hospitalization, increased healthcare costs, and elevated morbidity and mortality, underscoring the need for innovative therapeutic approaches that target both the pathogen and the host. To bridge the dynamic interplay between bacterial pathogens and the host immune system with emerging therapeutic innovations, this narrative review first examines the biological mechanisms underlying bacterial resistance. It then explores therapeutic strategies beyond conventional antibiotics, providing an overview of current approaches and their limitations, including drug repurposing, bacteriophage therapy, and CRISPR-Cas technology. The review subsequently focuses on antibacterial immunotherapy, discussing a broad range of emerging approaches, including probiotics, monoclonal antibodies, cell-based therapies, host-directed therapies, aptamers, nanotechnology-based platforms, cytokine-based therapies, and antimicrobial peptides. An integrated overview of preclinical evidence, clinical studies, and FDA-approved therapies is presented to assess the translational potential of immunotherapy strategies in combating AMR. Scientific, regulatory, manufacturing, and implementation challenges that influence their successful translation into clinical practice are discussed throughout. By integrating the biological basis of host-pathogen interactions with emerging antibacterial therapeutics and their translational development, this review provides a comprehensive framework for evaluating innovative strategies against antimicrobial resistance. In contrast to modality-focused reviews, it offers a unified perspective that highlights the complementary roles of pathogen-targeted and host-directed interventions and identifies future opportunities to improve the prevention and management of multidrug-resistant bacterial infections.

Emerging CRISPR-Cas, Antibody, and Nanomedicine Strategies Against Antimicrobial Resistance in Critical Care

Antimicrobial resistance (AMR) has become one of the defining threats to survival in intensive and neonatal intensive care units, where vulnerable hosts, invasive devices, and heavy empirical antibiotic exposure converge to select for multidrug- and extensively drug-resistant ESKAPE and non-aeruginosa Pseudomonas pathogens. We conducted a narrative-scoping synthesis of the peer-reviewed literature identified through targeted searches and, restricted to English-language articles addressing AMR mechanisms and next-generation, non-traditional therapeutics in critical care; reference lists were hand-searched, and findings were organized thematically rather than statistically pooled. Four convergent, non-traditional pillars emerged — programmable CRISPR-Cas and CRISPRi genomic tools, monoclonal antibodies and bioconjugates, antimicrobial and anticancer peptides (notably proline-rich peptides), and stimuli-responsive nanozymes and exosomal carriers — each capable of bypassing classical resistance mechanisms while, at least in preclinical models, sparing commensal microbiota. Persistent translational barriers include bedside diagnostic blindness to biofilm-embedded organisms, an unresolved neonatal and pediatric pharmacokinetic/pharmacodynamic void, and stewardship frameworks that still largely ignore the human resistome. Bridging bench-to-bedside gaps will require standardized biofilm models, dedicated pediatric PK/PD trials, artificial-intelligence-assisted therapeutic design, and microbiome-conscious stewardship, particularly given resource disparities across low- and middle-income settings.

Precision Medicine in Combating Antimicrobial Resistance: A Comprehensive Review.

Antimicrobial resistance (AMR) represents one of the most pressing threats to global public health, undermining the effectiveness of modern antimicrobial therapy and challenging decades of medical progress. This comprehensive review examines the transition from broad-spectrum empirical therapy toward precision medicine as an integrated framework for improving antimicrobial use and combating AMR. Precision medicine seeks to tailor treatment decisions by combining pathogen-specific genomic and resistance data with relevant host characteristics to optimize therapy while limiting unnecessary antimicrobial exposure and the selective pressures that drive resistance. The review synthesizes advances reported from 2020, highlighting established and emerging approaches including rapid molecular diagnostics, next-generation sequencing, CRISPR-based detection, machine learning (ML)-assisted decision support, precision dosing, and targeted therapeutics such as bacteriophage therapy, antimicrobial peptides, and bacterial proteolysis-targeting chimeras. Rather than functioning as isolated technologies, these approaches achieve their greatest clinical value when integrated within antimicrobial stewardship programs and a One Health framework that recognizes the interconnected human, animal, and environmental drivers of resistance. Despite considerable progress, important challenges remain, including equitable access to advanced technologies, interpretation of increasingly complex datasets, workforce and infrastructure limitations, and evolving regulatory pathways for novel diagnostics and therapeutics. This review concludes that while precision medicine is not a standalone solution, its successful implementation will depend on coordinated integration of diagnostics, host factors, computational tools, pharmacological optimization, and stewardship strategies to improve patient outcomes while preserving the long-term effectiveness of existing antimicrobials.

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.