Phage Delivery of CRISPR Interference System to Aid AMR Bacteria Sensitization

The spread of antimicrobial resistant (AMR) bacteria is a global problem that is taking a toll on both the health system and world economics. The process of new antibiotic development is slow, costly and unable to keep up with the insurgence of new AMR strains. New strategies and tools to fight this war need to be studied and developed to help get some of this challenge weight off the health system and try to tackle it from different angles. By the means of synthetic biology and the different aspects and tools that come with this discipline, the synthesis of new systems that can be employed to slow down the rise of this issue is possible. In this work we studied, characterized and exploited M13 bacteriophages as scaffolds to deliver a CRISPR interference (CRISPRi) system that achieved a partial re-sensitization of two different laboratory strains resistant to either meropenem or colistin. The AMR genes we targeted were blandm1 that aids resistance to meropenem, and mcr1 that aids resistance to colistin. Other than testing the system, we also investigated two different approaches for the evaluation of the results and the computing of the growth delay obtained by the circuit we employed. With this work we shine a light on the issue of AMR bacteria and investigate a new approach for their treatment with the hope that in the future it could be seen as a valuable alternative to help the classic antibiotic treatment.