The work builds on the results of microarray analysis and deals with the analysis of differential expression of muscle tissue in pigs. Using quantitative real-time RT-PCR the expression of 12 genes (ANXA1, ANXA2, BTG2, CSRP2, DAB2, EMP3, HBEGF, IL18, PRKAR1A, SGCD, SORT1, TGFB2) involved in the development and growth of muscle in pigs was validated. Differential expression of two muscles (m. longissimus lumborum et thoracis and m. biceps femoris) between Wild Boar and Czech Large White, which differ significantly in growth, was determined. All genes were overexpressed in Czech Large White. In order to resequencing and polymorphism detection, IL18 gene, encoding proinflammatory cytokine was selected. IL18 was previously associated with lipid metabolism, insulin-resistance and skeletal muscle function. In fragments covering exon 2, 4 and 5 a total of 40 polymorphisms were found. PCR-RFLP methodology was designed for two polymorphisms: ENSSSCG00000015037:g.11237G>A in exon 2 (MspI) and g.19480G>A in intron 5--6 (VspI) and the association analysis with the performance and meat quality traits in selected populations in pigs (n = 344 and 669) was evaluated. IL18 was associated with growth, muscle formation, fat deposition and meat quality traits. IL18 could be potentially useful for MAS in pigs.

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Diplomová práce se zabývá studiem genové regulace v Clostridium beijerinckii NRRL B-598, pro následné odvození genové regulační sítě bakterie C. beijerinckii NRRL B-598. V teoretické části této práce je uvedena obecná nomenklatura problematiky genové regulace se zaměřením na nomenklaturu genových regulačních sítí. Následně jsou zde popsané laboratorní metody, sloužící pro získání vhodných dat popisující expresi genů. Tato data jsou základem pro studium genové regulace a návrhy genových regulačních sítí. Práce se zaměřuje především na technologii RNA-Seq a stručný popis laboratorních dat získaných ze zmíněné bakterie C. beijerinckii NRRL B-598. V praktické části se práce zabývá předzpracováním těchto surových laboratorních dat a následným studiem genové regulace se zaměřením na odvození operonů a vytvoření prvních genových regulačních sítí pomocí různých přístupů pro C. beijerinckii NRRL B-598.

The aim of this study was to confirm that the polyunsaturated fatty acids n-3 (n-3 PUFA) have a positive effect on plasma lipids. These acids can reduce cholesterol by increasing gene expression Insig-1 while decreasing the expression of genes encoding Hmgcr and Ldlr. We tested in experimental rats, which were added to the feed mixture of 6 % safflower oil , 6 % fish oil or 6 % of the oil from the algae Schizochytrium. Relative gene expression was Insig-1 in the test group with addition of fish oil to 120% of controls (P<0.05) and in the group with addition of oils from algae Schizochytrium the relative expression of 170 % of control (P<0.05). These results confirm our hypothesis, only a part, as the relative expression of the gene and Hmgcr and Ldlr was in the test group with addition of fish oil 103% (P>0.05) and 101 % of control (P>0.05) and in the group with addition of oils from algae Schizochytrium the relative expression of 117% (P>0.05) and 156 % (P>0.05) compared to control. Thus, to reduce the relative expression of these genes did not. However, we have shown that n-3 PUFA contribute to a reduction in plasma cholesterol and in this case up to 20 % of control. The concentration of cholesterol in the group with addition of safflower oil was 1.35 mmol.l-1, the group with the addition of fish oil 0.98 mmol.l-1.

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Gene editing with the CRISPR/Cas9 system is one of the options that sets a new trend in the development of gene therapy. The most commonly used delivery of DNA into the cells are via viruses. Nevertheless, they are often unable to take CRISPR/Cas9 system, which can be bigger than several kb. Nanoparticles (NPs), as non-viral transporters, seem to be a good alternative delivery system. For this work magnetic Fe3O4 NPs (MNPs) were selected, because of their excellent properties such as multifunctionality, biocompatibility, easy degradation and simple synthesis. The aim of this work was to synthesise MNPs and a complex of MNPs coated with PEI/CRISPR-Cas9 plasmid and to characterize them by physicochemical methods. The created complex MNPs/PEI/CRISPR-Cas9 was defined by exact parameters that are suitable for possible cell uptake. The hypothesis of stabilization of the MNPs/CRISPR-Cas9 plasmid complex by polyethylenimine (PEI), which can also protect plasmid DNA against restriction endonucleases, was verified. Next a stable modified cell line HEK293-TLR3, designed to evaluate the efficacy of double strand break (DSB) repair by nonhomologous end joining (NHEJ) or homologous recombination (HR) was, transfected with the synthesised MNPs/PEI/CRISPR-Cas9 complex. The results indicate a 25% transfection...

(English) Phosphoinositides (PIs) are glycerophospholipids with a negative charge. As components of cell membranes, PIs are involved in membrane and cytoskeletal dynamics, cell movement and signalling, and the modulation of ion channels and transporters. Apart from the cytoplasm, phosphoinositides also localise to the cell nucleus. PIs play a role in crucial nuclear processes, such as DNA transcription, pre-rRNA and pre-mRNA processing, cell differentiation, DNA damage response, or apoptosis. Phosphatidylinositol 4-phosphate (PI(4)P) and phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) are the most abundant phosphoinositides in the cell. However, their exact localisation and function in the nucleus are largely unknown. Here, we describe their localisation at super-resolution level and their involvement in some nuclear processes. PI(4)P is present in nuclear lamina, nuclear speckles and nucleoli, and it forms small foci in nucleoplasm. The majority of nuclear PI(4)P localises to the nucleoplasm, whereas almost 16 % is present in nuclear speckles. On the other hand, the majority of nuclear PI(4,5)P2 localises to nuclear speckles, almost 30 % localises to nucleoplasm and the lesser portion to nucleoli. In the nucleoplasm, PI(4,5)P2 forms small foci called nuclear lipid islets (NLIs). Their core is...