Phd Degree / Doktora
Permanent URI for this collectionhttps://hdl.handle.net/11147/2869
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Doctoral Thesis Biochemical and Functional Characterization of Circular Rnas Differentially Expressed in Cisplatin-Treated Hela Cells(01. Izmir Institute of Technology, 2023) Akgül, Bünyamin; Akgül, Bünyamin; 04.03. Department of Molecular Biology and Genetics; 04. Faculty of Science; 01. Izmir Institute of TechnologyCircular RNAs (CircRNAs) are a novel class of single-stranded, covalently-closed RNA molecules. Functional investigations of the circRNAs provide insight into the mechanisms underlying gene regulation and cellular responses, which could ultimately lead to the development of new therapies for a wide range of diseases. In this thesis, four cisplatin (cis-diamminedichloroplatinum II, CP)-responsive circRNAs, circGALNT2, circBNC2, circBIRC6, and circCLASP1, were validated. The reverse genetics approaches, such as knockdown and overexpression strategies, showed that circCLASP1 is required for the proliferation of HeLa cells. The knockdown of circCLASP1 disrupts proliferation in HeLa, and its overexpression restores impaired proliferation. Further analyses revealed that circCLASP1 knockdown sensitizes HeLa cells against 20 μM and 40 μM cisplatin treatments. Interestingly, an IC50 dose of cisplatin causes Annexin V-/7AAD + cell death rather than apoptosis when combined with circCLASP1 knockdown. In light of these findings, five circRNA/miRNA/mRNA regulatory networks were constructed using computational approaches. Additionally, a transcriptomics analysis after circCLASP1 knockdown has supported all of these findings in that muscle cell proliferation genes were significantly altered upon circCLASP1 knockdown in HeLa cells. In conclusion, the findings suggest that the knockdown of circCLASP1 represses proliferation and sensitizes HeLa cells against cisplatin. CircCLASP1-knockdown mediated differential gene expression indicates proliferation, ROS response, iron metabolism, lipid peroxidation, and cell death. Further studies are needed to elucidate the precise mechanism of circCLASP1-mediated cell death and proliferation in muscle cells or liver cells and ROS-related diseases.Doctoral Thesis Elucidation of Boron Tolerance Mechanisms in Puccinellia Distans (jacp.) Parl. Using a Transcriptomic Approach(Izmir Institute of Technology, 2017) Öztürk, Saniye Elvan; Frary, Anne; Frary, Anne; 04.03. Department of Molecular Biology and Genetics; 04. Faculty of Science; 01. Izmir Institute of TechnologyThe amount of boron in soil is important for agronomic plants. As an abiotic stress condition, boron toxicity causes significant decreases in crop yields. Puccinellia distans (Jacq.) Parl. (P. distans), common alkali grass, is found throughout the world and can survive in soils with boron concentrations that are lethal for other plant species. Indeed, P. distans accumulates very high levels of this element. Despite these interesting features, very little research has been performed to elucidate the boron tolerance mechanism in this species. In this study, P. distans samples were analyzed by RNA sequencing to identify genes and miRNAs related to boron tolerance and hyperaccumulation. The results indicated that the hyperaccumulation mechanism of P. distans involves many transcriptomic changes including: alterations in the malate pathway, changes in cell wall components that may allow sequestration of excess boron without toxic effects, and increased expression of at least one putative boron transporter and two putative aquaporins. MiRNAs are also altered under stress conditions. The presence of miRNAs as stress regulator elements is an example of post-transcriptional regulation of stress related mechanisms. Additionally these small RNAs could affect their target genes by positive or negative regulation. Therefore, changes not only in miRNAs but also in their targets are important to understand their roles in hyperaccumulation. For example, downregulation of miRNA under stress could cause target accumulation. These mechanisms could be key in plant adaptation to new conditions. Elucidation of the boron accumulation mechanism is important in developing approaches for bioremediation of boron contaminated soils.