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Effect of ADAR1 enzyme on hepatitis C virus life cycle
Kubů, Martin ; Vopálenský, Václav (advisor) ; Fraiberk, Martin (referee)
Hepatitis C virus (HCV) is a virus of the family Flaviviridae whose genome consists of ,,+RNA" molecule. It causes the disease hepatitis Cepatitida typu C, which affects tens of millions of people worldwide. Although there is an effective treatment for this type of hepatitis, a preventive vaccine against the HCV virus has not yet been developed. Several ambigious works focused on the relationship between HCV and the enzyme adenosine deaminase acting on double-stranded RNA 1 (ADAR1) were published in the past. This dimeric double-stranded RNA binding enzyme is a part of innate immunity and causes catalytic conversion of adenosine to inosine, which is recognized by cellular mechanisms as guanine, which ultimately leads to mutations in the affected dsRNA molecule. The works published so far attribute an antiviral function to the ADAR1 enzyme in the context of HCV infection. However, vectors containing the entire HCV genome were not used in these works, and a cell line with deletion od the ADAR1 gene has never been used so far. The actual relationship between the ADAR1 enzyme and the HCV virus has not yet been reliably verified. The aim of this thesis was to gain a deeper understanding of the relationship between the HCV virus and the ADAR1 enzyme. For this task, HCV permissive Huh7.5 cell lines with...
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Sekvenování transkriptomu pro studium exprese genů u živočichů
Toufar, Jiří
In the introductory chapters, the thesis deals with the best description of the gene expression regulation process, which is an essential prerequisite for understanding the following chapters. Gene expression regulation covers regulation of transcription initiation, transcription elongation control, post-transcriptional modification and RNA editing. In the following chapters, the thesis deals with nucleic acid sequencing. Furthermore, classical sequencing methods, sequencing of the new generation, such as Illumina sequencing and the most modern third-generation sequencing methods such as SMRT-seq or Oxford Nanopore are also analyzed. Moreover, the thesis contains information on the comparison of selected methods. In the following sections, the thesis explains the term transcriptome and general procedure of the RNA sequencing experiment including single cell isolation, single-cell sequencing methods, and animal tissue sequencing. In the final chapters, the thesis explores the use of RNA sequencing and RNA-seq methods for specific purposes. In the conclusion, a comparison of RNA-seq with DNA/RNA microarrays was performed.
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Effect of adenosine deaminase acting on RNA on viral infection of eukaryotic cells
Kubů, Martin ; Vopálenský, Václav (advisor) ; Fraiberk, Martin (referee)
Double-stranded RNA is a molecule rarely found in a cell, but it is specific for viral infection. It is also a substrate of ADAR enzymes. These enzymes convert adenosin to inosine, which is recognized as guanosine by cellular machinery. Apart from editing activity, ADAR enzymes interact with cellular proteins, such as Dicer and protein kinase R, which together with editing affects viral replication. In this work, the information about antiviral activity of ADAR enzymes and their impact on infection of selected primarily human viruses is reviewed.
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