Julia Hettleman
Antibiotic resistance is an increasingly prevalent issue. My research uses CRISPR-Cas technology to identify and repress genes that contribute to antibiotic resistance to improve treatment of bacterial infections.
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Abstract:
Abstract:
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 identify the genes responsible for antibiotic resistance which can then be repressed to kill bacteria more effectively. To repress a gene, CRISPR uses small RNA strands (crRNAs) to find a location in the genome that matches the RNA sequence. Then, a CRISPR-associated protein (Cas) can bind to a site near a gene of interest and prevent transcription, allowing for varying degrees of gene expression. It is costly and time-consuming to synthesize crRNAs that match several genes in a genome, so CRISPR Adaptation-mediated Library Manufacturing (CALM) turns cells into “factories” that output a library of crRNAs. In this project, cells containing a crRNA targeting a gene of interest were grown in competition 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, or ability to thrive. For each gene tested, a low fitness suggests that the repression of that gene in combination with antibiotics more effectively kills bacteria. Using this data, drugs can be developed to repress the identified genes and used with antibiotics to treat bacterial infection more effectively.
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