Our human practice efforts in terms of engagement directly led to information that we incorporated into our project design.
Initially unsure of the best way to address the issue of antibiotic resistance via outreach, we performed extensive research on the subject. The publications we read mentioned many causes for the rapid pace of resistance development, including the overprescription of antibiotics in developed countries. We concluded that we needed the perspectives of clinicians if we were to understand why doctors overprescribe antibiotics, and how a group of undergraduates could hope to persuade doctors to prescribe antibiotics more appropriately.
As such, we interviewed Drs. Persell, Postelnick, and Scheetz, all of whom work on the research and clinical side of medicine through the Feinberg School of Medicine and Northwestern Medicine. These interviews revealed a major reason doctors prescribe unneeded antibiotics: patient demand. Instead of persuading doctors to change their behavior, we decided to educate the public on appropriate use and prescription of antibiotics.
In addition, the three physicians agreed that CRISPR would be more effective if targeted to knock out resistance genes for existing antibiotics rather than serving as a novel antibiotic by targeting other bacterial genes. Existing antibiotic resistance proteins, such as beta-lactamase and fluoroquinolones, are safer targets and result from well-characterized sequences. This advice incorporated well into the design of CRISPR Capsules—a realistic future aim would be to introduce Cas9 encapsulated in OMVs to a cell already containing a guide RNA for a selection marker such as ampicillin. If Cas9 enters the cell and cuts the resistance gene, it will not grow on that antibiotic. This would be a proof of concept for targeting antibiotic resistance with CRISPR, and also a simple test of successful OMV delivery.
One interview question asked the clinicians to identify what they considered the most worrying antibiotic-resistant infections. Their answers included hospital skin infections like Pseudomonas aeruginosa and Acinetobacter baumannii. This also informed our project design, because topical infections would be safe early applications of a bacterially-derived product to evaluate immune response to the therapy.