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Clinical classification of sequence variants in non-coding regulatory regions in breast cancer susceptibility genes.
Bubáková, Eliška
;
Ševčík, Jan
(advisor) ;
Vodička, Pavel
(referee)
Inactivation of tumor supressor gene BRCA1 causes a life-long risk of breast carcinoma development. Genetic screenings of indicated individuals from high-risk families help to identify large number of sequence variants in known predisposing genes. Majority of discovered variants doesn't have clinical significance yet which causes a big problem for diagnostics. Some of these variants are found within regulatory non-coding regions of gene. A part of the clinical classification of variants is their functional characterization. The goal of this thesis was to create a model system for functional characterization of variants in non-coding regions and to verify its function. Model system was based on targeted gene manipulation by co-transfecting CRISPR-Cas9 construct and donor construct that contained a portion of BRCA1 gene sequence with analyzed modifications, into U2 OS cells. The cells have stably integrated DR-GFP system which allows the activity of homologous recombination (HR) to be determined. Monoallelic modifications were induced into U2 OS cells. These modifications were in a Kozak sequence region of BRCA1 gene. Expression level of BRCA1 mRNA was determined by qRT-PCR, which showed the same levels of mRNA in all cells with analyzed alterations. Next, expression level of BRCA1 protein was...
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Interactions of metal nanoparticles with microorganisms
Bubáková, Eliška
;
Beranová, Jana
(advisor) ;
Zikánová, Blanka
(referee)
This thesis deals with metal nanoparticles and their antibacterial effects. Nanotechnologies have been paid intensive attention both in scientific and public area. Some metal nanoparticles possess provable antibacterial potential. Nowadays, when the growing resistance of pathogenic bacteria represents an important issue, metal nanoparticles could be one of the potential solutions to the problem. Interactions of metal nanoparticles with bacteria have however not yet been fully understood. It is also evident that their effects depend on various physical and biological factors. The cytotoxicity of nanoparticles to eukaryotic organisms has not been explored in depth either. These all are the reasons why the practical use of nanoparticles has still many limitations. This thesis summarizes information available on the topic of bactericidal effects of metal nanoparticles.
Detailed record
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