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3rd CCP Phenogenomics Conference 2021: Abstract Book
Sedláček, Radislav
The Conference was divided into two blocks. The first day was devoted to the theme “Human diseases and models“. The second day was specifically dedicated to preclinical development, including covid-19, which focused on translation of the basic research into the application. The Conference provided an excellent opportunity to support networking and interactions among the CCP users and experts.
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2nd CCP Phenogenomics Conference 2020: Abstract Book
Sedláček, Radislav
The Conference was divided into two blocks. The first day was devoted to the theme “Human diseases and models“. The second day was specifically dedicated to preclinical development, including covid-19, which focused on translation of the basic research into the application. The Conference provided an excellent opportunity to support networking and interactions among the CCP users and experts.
Plný tet:  PDF
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Genome editing using programmable endonucleases
Hanečková, Radmila ; Sedláček, Radislav (advisor) ; Sýkora, Michal (referee)
Programmable endonucleases are engineered proteins that recognize specific nucleotide sequences and that are capable of introducing double-strand breaks within these sequences. Zinc-finger nucleases have been used extensively as a tool in genome editing, the practice of introducing changes into genomes of cell lines or whole organisms as a way to study gene function. Recently, new types of programmable endonucleases have emerged in the form of transcription activator like effector (TALE) nucleases and the CRISPR/Cas system. The types differ in respect to their mechanism of function, accessibility, selectivity, frequency of off-target cleavage and cytotoxic effects. Here, we compare zinc-finger nucleases, TALENs and the CRISPR/Cas system and explore their current and possible future applications in a broad spectrum of research ranging from developing genetically modified organisms to gene therapy. Powered by TCPDF (www.tcpdf.org)
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Influence of extracellular matrix environment on gene expression in liver myofibriloblasts
Jiroutová, Alena ; Kanta, Jiří (advisor) ; Ehrmann, Jiří (referee) ; Sedláček, Radislav (referee)
Influence of extracellular matrix environment on gene expression in liver myofibroblast (summary) Hepatic stellate cells (HSC) and liver myofibroblasts (MF) are two cell populations most likely responsible for the synthesis of majority of connective tissue components in fibrotic liver. They differ in their origin and location in the liver, and in the spectrum of genes they express. HSC are located in Disse spaces of normal rat liver around the sinusoids, in fibrotic liver they become activated, proliferate and they undergo transdifferentiation into myofibroblast-like cells. Myofibroblasts are heterogenous cell population that consists at least of portal pMF, septal sMF and interface iMF. pMF, which are adjacent to bile duct epithelia, may be a mediator of billiary type fibrosis. sMF are located within and along the collagenous septum in cirrhotic liver. Little is known about the expression of genes involved in connective tissue metabolism in MF cultured in fibrin or collagen gels that more closely resemble natural cell environment. Fibrin is deposited in liver at sites of injury and collagen type I forms a substantial part of fibrotic septa. In our study oligo cDNA array analysis was used to determine gene expression in quiescent HSC, activated HSC and MF isolated from both normal and CCl4-cirrhotic liver....
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Postranslation modifications affecting function of nuclear localization signal
Šebrle, Erik ; Sedláček, Radislav (advisor) ; Venit, Tomáš (referee)
Transport of proteins to the nucleus through a nuclear envelope is controlled mostly via nuclear localization signal (NLS). Nuclear localization signal is rich in positively charged amino acids arginine and lysine. It was observed that activity of this NLS could be regulated through a phosphorylation of serine in its close proximity. Either a phosphorylation of serine or phosphomimetic changes of these "presequences" could represent an important mechanism regulating a localization of protein in cells in relation to a cellular activation. In our laboratory was identified protein - Fragile X mental retardation syndrome 1 neighbor (Fmr1nb), whose cellular localization could be driven by this posttranslational modification.
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Generation and analysis of mutant mouse models to study pathophysiological roles of KLK5 and KLK7 in epidermis
Kašpárek, Petr ; Sedláček, Radislav (advisor) ; Stopka, Pavel (referee) ; Machoň, Ondřej (referee)
Kallikrein-related peptidases (KLKs) constitute a family of closely related serine proteases encoded by genes clustered in one chromosomal locus. KLKs are widely expressed in a variety of tissues and numerous in vitro experiments suggest their important roles in many physiological and pathological processes. However, the biological roles of KLKs in vivo are often obscured mainly due to unavailability of suitable animal models. Although gene deficient mouse models were generated for several KLK genes, they had limited use for understanding the roles of individual proteases in the complex environment in vivo. One of the main obstacles which hampers in vivo analysis is partial functional overlap between some KLKs. This makes traditional single-gene deficient animal models an inadequate tool to address the biological impact of the gene deficiency as compensatory mechanisms often result in a lack of phenotype. In this work, we used the transcription activator-like effector nuclease (TALEN) technology to generate several novel mutant mouse models to study the complex KLK proteolytic pathways and their roles in healthy organism and in disease. We prepared a novel mouse model for Netherton syndrome (NS), an autosomal recessive skin disorder caused by mutation in the gene SPINK5, which encodes the KLK-inhibitor...
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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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