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Knockout isoforem metalothioneinu pomocí metody CRISPR-Cas9 u adherentních tkáňových kultur
Duda, Jakub
It is not too long ago, that genome modifications were very complex, or, in some cases, almost impossible to achieve, but with the discovery and description of the CRISPR system in prokaryotes, a lot of breakthroughs came to the human knowledge, and thanks to some of these breakthroughs the CRISPR system has been modified to be used as a genome editing tool in eukaryotic organisms. Genetic modifications could in theory be the key to cancer therapy in modern pharmacy, and this thesis focuses on Metalmetallothionein (MT) proteins, which are commonly found in organisms and are beneficial to the organism (as inhibotors of oxidative stress, or as antioxidants binding heavy metals, for example), but can also be dangerous. Because, according to the isoform, and also the type of tissue, MTs can cause the development of cancer tissue and tumour growth, sometimes because of their presence, in other cases because of their absence. This thesis was focusing on the possible knock-out of all isoforms of MT at once, using CRISPR/Cas9 method, and thereby infucencing one of the very important factors in carcinogenesis. The result of this thesis is the achievement of knock-out of the MT3 isoform.
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