Engineering Red Light-Activated Guanylate Cyclase for Use in Pathogen-Host Interactions
presentationposted on 17.11.2014, 00:00 by Jesse Hinshaw
Host-pathogen studies are the method through which knowledge concerning how pathogens interact with their hosts is obtained. Currently many of these studies are performed by knocking-out genes in pathogens and then infecting mice with the genetically modified pathogen. This method of knocking-out genes is very imprecise though, as it is difficult to control the temporal aspects of infection such as toxin production during specific stages of infection. My project aims to produce a new and more effective method for studying how pathogens interact with hosts via creating a near infrared (NIR) light-activated enzyme that produces a secondary messenger, in this case the enzyme is a guanylate cyclase and the messenger is cGMP (cyclic guanosine mono-phosphate). The light activated portion of said enzyme is a bacteriophytochrome sensitive to NIR light that deeply pierces mammalian tissue. By being able control the activation of a guanylate cyclase via NIR light, researchers will be able to regulate the production of cGMP and therefore expression of genes via a transcription activator that senses cGMP in spatially and temporally accurate manner. My project focuses on producing DNA encoded for such a NIR light-activated cyclase via two methods, fusion of a known guanylate cyclase domain to a bacteriophytochrome domain and a series of point mutations in a known adenylate cyclase to produce a guanylate cyclase. This project, when completed, will yield a new tool in the arsenal of researchers around the world hoping to develop new methods to learn about and combat pathogens.