New York-Metro Synergistic Interactions Between Essential Genes and Antibiotics in Staphylococcus Aureus

Overview

Antibiotic resistance is an increasingly prevalent issue as bacteria adapt to the overprescription of antibiotics, making the treatment of bacterial infections more difficult. To combat this, the novel gene-editing technology, CRISPR, can be used to assess interactions between certain genes and traditional antibiotics in a bacterial genome. CRISPR Adaptation-mediated Library Manufacturing (CALM) is a method of manufacturing a library of crRNAs - a key molecule in CRISPR’s targeting mechanism - allowing for gene repression via CRISPR inhibition (CRISPRi) of up to 90% of genes. In this project, CALM -CRISPRi libraries were constructed and treated with antibiotics. The libraries were then sequenced to identify which gene perturbations reduced bacterial fitness in the presence of antibiotics. To validate the screening, cells containing a replicated crRNA targeting a gene of interest were grown in co mpetition with common competitor cells lacking crRNAs. The cells were grown in the presence of various antibiotics to identify any synergy between the antibiotics and the gene repression. DNA was extracted from these populations and analyzed, resulting in a final calculation of the cells’ fitness. We identified synergy between similar processes; for example, the repression of a cell wall synthesis gene synergized with an antibiotic targeting the cell wall. Additionally, simultaneous repression of cell wall/cell membrane and DNA synthesis/replication genes revealed widespread synergy between the two processes. Using this data, drugs can be developed to repress the identified genes and used with antibiotics to treat bacterial infection more effectively.

Competition history

• JSHS 2025 Category not listed

Resources

• National JSHS abstract book ↗

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