The rapid rise of multidrug-resistant (MDR) superbugs and the declining antibiotic pipeline are serious challenges to global health. Rational design of therapeutics can accelerate development of effective therapies against MDR bacteria. In this talk, I will describe multi-pronged systems, synthetic biology, and nanobiotechnology based approaches being devised in our lab to rationally engineer therapeutics that can overcome antimicrobial resistance. Our lab uses Systems biology approach by studying the transcriptome of bacteria exposed to antimicrobials, or extreme conditions such as microgravity during space flight, or quantum dots, to identify mechanisms by which bacteria can enter an “adaptive resistance” state. Our analysis has uncovered that bacteria can sample a dynamic gene regulatory space that can result in multiple solutions to the “same stress” problem; while gene expression levels can be highly variable in adapted populations, we find reproducible shifts in gene expression variability provide insights into key genes involved in adaptive resistance. Based on these insights we have developed Synthetic biology-based approach dubbed “Controlled Hindrance of Adaptation of OrganismS” or “CHAOS” to slow the evolution of antibiotic resistance by interfering with processes involved in adaptive resistance. Using CRISPR based technology, we rationally engineer library of synthetic genetic devices for multiplexed activation and inhibition of native gene expression of key essential and stress-response gene networks. Here we show that CHAOS approach leads to predominant negative epistasis with severe loss of fitness during adaptation to a range of toxins, including disinfectants and antibiotics, and eventual “slowing” down of bacteria's ability to adapt. To translate our findings into the clinical setting, we engineer antisense therapeutics that can block translation of any desired gene in a pathogen-specific manner for targeted inhibition. Using this approach we are building a Facile Accelerated Specific Therapeutic (FAST) platform for the accelerated development of novel antibiotics against MDR bacterial clinical isolates as well as any emergent bacterial threats. The FAST approach can also be used to decrease or increase expression of desired genes in mammalian cells for identifying key players of disease phenotypes. Finally, I will also present a nanobiotechnology based approach involving development of a unique semiconductor material based quantum dot-antibiotic (QD ABx) which, when activated by stimuli, release reactive oxygen species to eliminate a broad range of MDR bacterial clinical isolates including methicillin-resistant Staphylococcus aureus, extended-spectrum β-lactamase producing Klebsiella pneumoniae and Salmonella typhimurium, and carbapenem-resistant Escherichia coli. The CHAOS, FAST and QD Abx platforms and inter-disciplinary approaches presented in this talk offer novel methods for rationally engineering new therapeutics to combat disease challenges.