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Generation of conditional animal mutants to study gene function in vivo
Herrmannová, Pavlína ; Sedláček, Radislav (advisor) ; Novák, Josef (referee)
Conditional gene targeting allows spatial and temporal control of genetic modifications and is used to study gene functions in specific tissues or cell types. Gene targeting may lead to inactivation of the gene by insertions or deletions. Conditional gene targeting uses various methods for generation of transgenic mutant animals, such as technology of targeted disruption of gene using embryonic stem cells, methodology based on bacterial artificial chromosomes, or a new revolutionary technology of targeted disruption of genes using programmable nucleases, which is rapidly evolving and seems to be more efficient and cheaper method for conditional gene targeting. The aim of this work is to overview methods and technologies for generation conditional animal models, and overview conditional recombination systems with emphasis on inducible systems, and also provides a summary of the main international resources for rodent mutagenesis. Key words: transgenic animal model, gene, targeting, conditional allele
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Investigation of proteolytic enzymes expression in different tissues at the transgenic animal model of Huntington disease by means of biochemical and immunohistochemical methods
Kocurová, Gabriela ; Dršata, Jaroslav (advisor) ; Novotná, Eva (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Gabriela Kocurová Supervisor: Prof. MUDr. Jaroslav Dršata, CSc. Title of diploma thesis: Investigation of proteolytic enzymes expression in different tissues at the transgenic animal model of Huntington's disease by means of biochemical and immunohistochemical methods Background: Huntington's disease (HD) is a neurodegenerative disorder that is caused by an expansion of a polyglutamine (polyQ) domain in the huntingtin (Htt) protein. Because it is known that mutant Htt and especially its small proteolytic fragments are toxic to neurons (particularly those in the striatum and cortex), it has been suggested that proteolysis of mutant huntingtin (mHtt) might play an important role in HD pathogenesis. Therefore, the aim of the present study was to examine the expression of endogenous and mtHtt and possible participation of the proteolytic enzymes from the group of caspases, matrix metalloproteinases (MMPs), kallikreins (KLKs) and calpains in HD pathology of brain tissue. Methods: In this study we used WT and TgHD minipigs for N-terminal part of the human mtHtt (548aaHTT-145Q, both F2 generation, age 36 months; F3 generation, age 48 months in additional experiment), R6/2 mice were used as...
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Generation of conditional animal mutants to study gene function in vivo
Herrmannová, Pavlína ; Sedláček, Radislav (advisor) ; Novák, Josef (referee)
Conditional gene targeting allows spatial and temporal control of genetic modifications and is used to study gene functions in specific tissues or cell types. Gene targeting may lead to inactivation of the gene by insertions or deletions. Conditional gene targeting uses various methods for generation of transgenic mutant animals, such as technology of targeted disruption of gene using embryonic stem cells, methodology based on bacterial artificial chromosomes, or a new revolutionary technology of targeted disruption of genes using programmable nucleases, which is rapidly evolving and seems to be more efficient and cheaper method for conditional gene targeting. The aim of this work is to overview methods and technologies for generation conditional animal models, and overview conditional recombination systems with emphasis on inducible systems, and also provides a summary of the main international resources for rodent mutagenesis. Key words: transgenic animal model, gene, targeting, conditional allele
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