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Explain The Mechanism Of RIG-I Recognizes Bacterial Ligands In Glial Cells?

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RIG-I recognizes bacterial ligands in glial cells. RIG-I is traditionally viewed as a viral pattern recognition receptor, however, more recently the pathogen ligands for RIG-I have been expanded to include bacteria nucleic acids. In non-CNS cell types RIG-I has been documented to identify bacteria RNA directly or DNA indirectly via an RNA polymerase III dependent mechanisms. RIG-I dependent recognition of bacterial nucleic acids was demonstrated to be pathogen and cell type specific. In glial cells the role and mechanism of RIG-I in pathogen detection has not fully been explored. We have previously demonstrated expression of RIG-I and the contribution of RIG-I to recognition of RNA and DNA viruses. However, the requirement of RIG-I for …show more content…

Confocal imaging of RIG-I recognition of bacterial ligands and nano-TNAs. In this proposal we will investigate the subcellular localization of bacteria and bacterial nucleic acids in relation to the cytosolic innate immune receptor, RIG-I. In order to accomplish this goal we will combine traditional immunofluorescence for RIG-I and cellular compartment protein markers with the use of commercially available products EU and EdU to label bacteria RNA and DNA, respectively. The Thermo Fisher Scientific products EU and EdU were originally developed to detect global RNA transcription and measure de novo DNA systhesis or S-phase synthesis, respectively. By growing bacteria in the presence of these products prior to infection of glial cells we can track not only bacteria but nucleic acid subcellular localization. This will provide insight into the accessibility of nucleic acids to host innate immune receptors. By innovatively combining immunofluorescence and nucleic acid labeling we will characterizing the localization of bacteria, bacterial nucleic acids, and RIG-I over a given time course providing valuable insight into the mechanisms of host cell recognition of bacteria in glial cells.
Nano-TNAs present significant advantages over traditional counterparts for therapeutic and adjuvant applications.

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