Regulation of Interleukin-6 by an Upstream Open Reading Frame

Claire Kelly*, Savannah Sims, and Gordon Meares

Department of Microbiology, Immunology, and Cell Biology, West Virginia University, Morgantown, WV, 26506

Presentation Category: Oral-Science & Technology (Oral Presentation #24)

Student’s Major: Immunology and Medical Microbiology

Interleukin-6 (IL-6) is a cytokine involved in inflammatory responses and is commonly increased in chronic diseases. This cytokine stimulates the synthesis of proteins, such as other cytokines and chemokines, involved in the inflammatory response and the growth and support of some immune cells. IL-6 is typically only expressed during inflammation to ensure proper control of the immune response. Regulation that controls this pattern of expression is not completely understood. Because the code to produce a protein must be transcribed from DNA to mRNA and then translated, there are a variety of potential mechanisms that could be controlling the expression of IL-6. We hypothesize that a key regulatory component may lie within the untranslated region of the mRNA transcript. To test this possibility, we used the CRISPR/Cas9 gene editing technique to create IL-6 human glioma cell lines containing different mutations in the upstream open reading frame (ORF). We then isolated and sequenced the IL-6 gene in our mutants to determine what mutations were introduced. We performed an enzyme-linked immunosorbent assay (ELISA) to measure the amount of IL-6 protein produced by each of these mutants under conditions that provoke IL-6 expression. We also measured the relative amount of the IL-6 transcripts (mRNA) produced under these conditions. Identifying the mechanism behind IL-6 regulation will allow us to better understand the neuroinflammatory response and the potential role in neurological diseases such as Multiple Sclerosis, Alzheimer’s disease, and ischemic stroke.

Funding: IMMB Internship

Program/mechanism supporting research/creative efforts: Other, Immunology and Medical Microbiology Internship