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National Repository of Grey Literature 2 records found  Search took 0.00 seconds. 
Directed evolution of mouse polyomavirus
 Váňová, Jana ; Španielová, Hana (advisor) ; Mašek, Tomáš (referee)
The method of directed evolution represents a new approach to generate proteins with new or altered properties. The principle of directed evolution is random mutagenesis of the coding sequence for a protein of our interest followed by selection of generated mutants for the desired property. The aim of this pilot study was to investigate the possibility of utilization of directed evolution for alteration of mouse polyomavirus original tropism and virus retargeting to a model prostate cancer cell line. To generate randomly mutated gene encoding the major capsid protein of mouse polyomavirus, which is responsible for the interaction of the virus with cellular receptor for viral cell entry, error-prone PCR and DNA shuffling methods were used. Production of viruses composed of mutant major capsid protein was ensured by Cre/loxP site-specific recombination. The thesis also dealt with the design and characterization of the system for viral mutant selection. It was found that the prostate cancer cell lines markedly vary in their ability to bind and internalize particles derived from mouse polyomavirus. This knowledge can be used for the preparation of virus-like particles for prostate cancer diagnostics in the future. The study demonstrated that the method of directed evolution can be used for production...
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Retargeting of viral particles for directed cargo delivery into cells
 Váňová, Jana ; Španielová, Hana (advisor) ; Kuthan, Martin (referee)
Due to their structure and ability to enter cells where they release their content, viral particles represent an attractive tool to deliver cargo to a cell. For therapeutic usage of viruses it is necessary to ensure the specific and highly efficient entry to target cells. This thesis offers an overview of methods used for virus retargeting with the intention to evaluate the success of retargeting in terms of specificity and efficiency of designed viral particles when entering a cell. On the basis of published data and considering the mechanisms of viral infection, the thesis demonstrates the difficulty to prevent the nonspecific viral particles from entering the cell and concludes that reaching total specificity is apparently impossible. Despite these small limitations, viral nanoparticles are a revolutionary therapeutic tool for delivering cargo to the cell and it is necessary to exploit their potential.
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