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Project Description

Content

Treating infectious diseases of the gastrointestinal (GI) tract with antibiotics disrupts a patient's gut microbiota and can increase the prevalence of antibiotic resistant strains. The increasing population of multi-drug resistant bacterial strains, both within and outside of health centers, has resulted in numerous infections which are becoming progressively difficult to treat. Additionally, it is a well-recognized fact within the global health community that traditional antibiotics do not represent a sustainable method of treatment for bacterial infections. There is a clear drive towards effective minimally invasive, prophylactic therapies for such ailments, but is a demand that so far, has not been adequately met. More specifically, therapies with high specificities for disease etiologies and minimal cross-talk among healthy tissues, organs, or symbiotic organisms are a highly desirable quality for treating infections. We envision a new paradigm for treating infections of the human gastrointestinal tract through exploitation of engineered probiotics that produce anti-microbials with high specificity for pathogens. The anti-microbials we are exploring do not utilize a one-fit all therapy mold, but target unique features specific to organisms at the genetic level. Towards this aim, we have utilized a general-purpose system that will be delivered to pathogenic bacteria from an engineered bacterial species found in the GI tract (Escherichia coli), which will cleave pathogenic genes at precise locations with single nucleotide resolution. To achieve specific genome targeting, we will utilize the CRISPR/Cas9 system with gRNA engineered to recognize genes from infectious bacteria that contribute to pathogenicity in humans. Our CRISPR/Cas9 system will be delivered from the engineered E. coli to infectious bacteria using bacterial specific phages, minimizing any side-effects to native microbiota and human-host cells. As a proof-of-principle for our engineered probiotic, we are starting by targeting Vibrio cholera, however we hope to expand the system to other pathogens of the GI tract.

References

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1. Overall project summary
2. Project Details
3. Materials and Methods
4. The Experiments
5. Results
6. Data analysis
7. Conclusions

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