Investigation of the effect of circadian rhythm on vaccine-acquired immune response with CRİSPR/CAS9 technique

Circadian rhythms are biological processes that occur through the regulation of gene expression over time. These processes influence many biological processes such as the sleep and wake cycle, hormonal release, metabolism and immune functions. Circadian rhythms are therefore important for the regular expression of genes and the proper regulation of the body. Since TIMELESS, a circadian gene, is also involved in various biological processes such as cell cycle and DNA damage response, it constitutes a broad research area. In this thesis, the relationship between the TIMELESS circadian gene and the immune system was investigated. For this purpose, firstly, the physicochemical properties, subcellular localization and 3D protein structure of TIMELESS were examined using in silico methods. Then, the evolutionary relationships of TIMELESS were determined by phylogenetic tree and motif analysis. Then, 52 miRNAs targeting TIMELESS were identified by in silico methods. These miRNAs were bioinformatically identified to be associated with cancer, immunity and infection. In addition, using the CRISPR/Cas9 method, the TIMELESS gene was knocked out in HEK293 cells and the expression levels of 84 innate and adaptive genes were examined by BCG vaccination. As a result of the findings, it was determined that the TIMELESS gene is associated with immune system functions. This thesis study may help to better understand the functions of the TIMELESS gene and other circadian genes, to better investigate the effects of circadian rhythms on health and disease, and to develop therapeutic approaches.