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Generation and analysis of double deficient transgenic mice for kallikrein-related peptidase 5 and kallikrein-related peptidase 14
Hanečková, Radmila ; Sedláček, Radislav (advisor) ; Fulková, Helena (referee)
Kallikrein-related peptidases (KLKs) constitute a highly conserved serine protease family. Based on in vitro experiments, KLKs are predicted to play an important role in a number of physiolog- ical and pathophysiological processes. However, their role in vivo remains not fully understood, partially due to a lack of suitable animal models. In this work, we aim to prepare a KLK5 and KLK14 double-deficient mouse model. Both KLK5 and KLK14 were proposed to be involved in epidermal proteolytic networks critical for maintaining skin homeostasis. However, both KLK5 and KLK14 single-deficient mouse models show minimal or no phenotype, likely due to similar substrate specificity resulting in functional compensation. Double-deficient mice cannot be easily obtained by crossing due to localization of the Klk5 and Klk14 genes within the same locus on chromosome 7. We report that KLK5 and KLK14 double-deficient mice were success- fully generated, mediated by transcription activator-like effector nucleases (TALENs) targeting Klk14 by microinjection of TALEN mRNA into KLK5-deficient zygotes. Furthermore, we show that KLK5 and KLK14 double-deficient mice are viable and fertile. We believe that these novel mouse models may serve as a useful experimental tool to study KLK5 and KLK14 in vivo.
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Statically indeterminate rod systems
Sedláček, Radek ; Řehák, Kamil (referee) ; Florian, Zdeněk (advisor)
The bachelors thesis is focused on methods of solving rod systems and that mainly statically indeterminate. At first is mentioned theoretical basis of rod systems, next description of selected methods for solving statically determinate rod systems and then description of methods for solving statically indeterminate rod systems which are demonstrated on model examples. The purpose of this thesis is to consider the possibility of using methods of solving statically indeterminate rod systems nowadays, which is devoted conclusion.
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Communication Server Design and Implementation
Sedláček, Robert ; Kenyeres, Martin (referee) ; Škorpil, Vladislav (advisor)
In the first part of this work, there is analysis of the industrial proprietary protocol S7 by Siemens. The analysis is performed by Wireshark between the programmable controller S7-1200 and Simatic NET OPC server. This protocol is based on other protocols whose proposals and solutions are completely or at least partially described in this work. On the basis of this information is created a communication library in C ++, which allows the transmission of variables. This library is subsequently used in the implementation of the communication server through which one can easily access variables in the programmable machine with a text protocol.
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Detached house
Sedláček, Radek ; Pospíšil, Jaroslav (referee) ; Kolář, Radim (advisor)
The subject of this thesis is a design of new building detached house with attic flat in Velké Karlovice. House has three flats and house is designed for ten to twelve permanent residents. The object is situated on a slightly sloping land on the outskirts of the village Velké Karlovice. Part of the building is a garage for three cars. The main entrance is oriented towards the main street. Plan dimensions of the house are 9,5 × 25 m. Vertical constructions are made of ceramic bricks of Porotherm, ceiling construction is designed using technology of Spiroll. Stairs construction is a precast element. The building is roofed by saddle roof with purlins. Part of the thesis is a project documentation for a construction including constructional details, fire safety, thermal engineering and acoustic solutions. The project documentation had been eleborated in an AutoCAD program.
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Newly-built of boarding house in Velke Karlovice
Sedláček, Radek ; Nováková, Eva (referee) ; Kolář, Radim (advisor)
The subject of this thesis is a design of new building boarding house with attic flat in Velké Karlovice. House has eight flats and house is designed for temporary accommodation, the capacity is maximum 20 guests. The object is situated on a large and almost a flat land at parcel no. 3640/1. Pension is detached building. The main entrance and arrival is oriented in the northern part of the building. Plan dimensions of the house are 9,5×15 m and 10×15 m. Vertical constructions are made of ceramic bricks of Porotherm, ceiling construction is designed using technology of Spiroll. Stairs construction is a precast element. The building is roofed by saddle roof with purlins.
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Creating a computational model for stress-strain analysis of parts of generator
Sedláček, Radek ; Fučík, Karel (referee) ; Návrat, Tomáš (advisor)
This thesis focuses on creating of a computational model for stress-strain analysis of parts of a generator. Computational model is parametric and it allows use of various geometric variants of the shaft and rotor lamination. Parametric computational model in APLD language is compiled in the attached sheet of software MS Excel after entering input values. After solving in software ANSYS 14.5 are results imported into MS Excel sheet for further post processing.
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Functional analysis of posttranscriptional gene regulation by TENT5A in biomineralization and metabolism
Aranaz Novaliches, Goretti ; Sedláček, Radislav (advisor) ; Hovořáková, Mária (referee) ; Tencerová, Michaela (referee)
Non-canonical poly-A polymerases, such as TENT5A, belong to the Terminal nucleotidyl transferases (TENTs) family and are crucial for mRNA protection, stability, and translation. A Tent5a knock-out (KO) mouse model was generated in our laboratory, which exhibited a phenotype in teeth, skeleton structure, and metabolism. In my PhD project, I aimed to characterize the molecular mechanism underlying these phenotypes and explore their potential connection to rare human diseases. I focused on the biological function of Tent5a gene in enamel development (amelogenesis) and mRNA stabilization. Micro-computed tomography and scanning electron microscopy revealed that Tent5a KO mice displayed thin, hypomineralized enamel with disrupted microstructure, a condition known as Amelogenesis imperfecta. Direct mRNA sequencing demonstrated that TENT5A is responsible for polyadenylation of amelogenin (AmelX) and other secreted proteins, leading to a shortened poly-A tail in Tent5a KO ameloblasts. Moreover, Tent5a KO mice disclosed impaired self- assembly of enamel matrix proteins (EMPs) such as AMELX and ameloblastin (AMBN), leading to compromised hydroxyapatite deposition and enamel formation. In addition to its role in teeth, I investigated the physiological functions of EMPs in other tissues, considering that EMP...
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4th CCP Phenogenomics Conference abstract book
Sedláček, Radislav
The fourth CCP Phenogenomics Conference was held as a hybrid meeting. The scientific committee selected the topic of rare diseases: experimental models & delivery of therapies as the main thematic focus of the 2022 Conference. The Conference provided again an excellent opportunity to support networking and interactions among the researchers, CCP staff, users and experts from the commercial sector.
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Mouse models for Angelman syndrome: generation and characterization
Syding, Linn Amanda ; Sedláček, Radislav (advisor) ; Vyklický, Ladislav (referee) ; Valeš, Karel (referee)
Angelman syndrome (AS) is a neurodevelopmental disease found in 1 to 10,000 to 40,000 births, exhibiting an equal gender ratio. Key characteristics of the disease include an ataxic gait with tremor, severe mental retardation, profound speech impairment and seizures. Behavioral deficits such as increased anxiety and autism spectrum disorder features is found in affected individuals as well. The disease stems from the imprinted region 15q11.2-13q where genes are either maternally or paternally expressed as a result of parent-of-origin specific expression of the alleles. There are four main genetic etiologies causing AS namely, i) a large deletion ranging from 4-6 Mb on the maternally inherited allele including imprinted and bi-allelically expressed genes, ii) maternal deletion of the Ubitiquin ligase E3 (UBE3A) gene, iii) paternal uniparental disomy and iv) imprinting defect leading to inappropriate methylation of the locus. So far, there is no cure for AS rather the symptoms are ameliorated using a multidisciplinary approach. The goal of the doctoral study was to further decipher the role of Ube3a and Gabra5 using two mouse models to gain more knowledge about the involvement of these two genes for future therapeutic interventions in for Angelman syndrome. One model generated was a full gene deletion...
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