Team:CU-Boulder

From 2014.igem.org

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CRISPR-Cas Mediated Phage Therapy

the Future Antibacterial Agent

University of Colorado at Boulder

Issue

In the last decade the number of antibiotic resistant bacteria has grown at a shocking rate, making these “superbugs” one of our top global health threats. This trend indicates that the golden age of antibiotics is ending and makes the search for new antibacterial treatments more important than ever. New bacterial treatments need the ability to adapt faster than the harmful bacteria that they target. Additionally, new treatments must preserve essential bacteria of the microbiome and specifically target pathogenic bacteria in a mixed population.

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Technology

New technology has recently emerged that allows us to achieve these goals. CRISPR-Cas9 consists of an endonuclease (Cas9) that is guided to a specific sequence within a genome by a CRISPR RNA component. Once targeted to the genome, Cas9 causes a double stranded break, killing the host bacterial cell. Because the killing relies on the sequence of the CRISPR guide RNA, which can be engineered to contain a short specific nucleotide sequence, unique genes within any pathogenic bacteria can be targeted.

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Our Project

Utilizing a non-replicating phage as a vector, we can efectively deliver the CRISPR-Cas9 machinery to cells and kill bacteria through CRISPR-Cas9–targeting of the genome. Ongoing experiments are aimed at demonstrating that sequence-specific killing can occur in a mixed population of cells as well as in an in vivo model. We believe that this technology, which we refer to as “Pathogen Assassin” will have broad applications ranging from medicine to industry and beyond.