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Preparation of STAT3-knockout glioblastoma multiforme cell line using CRISPR-Cas9 editing
Papinčáková, Dominika ; Skálová, Lenka (advisor) ; Laštovičková, Lenka (referee)
Charles University Fakulty of Pharmacy in Hradec Králove Department of Biochemical Sciences Candidate: Bc. Dominika Papinčáková Supervisor: prof. RNDr. Lenka Skálová, Ph.D. Title of diploma thesis: Preparation of STAT3-knockout glioblastoma multiforme cell line using CRISPR-Cas9 editing Glioblastoma multiforme (GBM) is one of the most common primary brain tumours in adults and one of the most fatal cancers in humans A key oncogene in GBM development and progression is the signal transducer and activator of transcription 3 (STAT3). It is one of the mediators of tumorigenesis, plays a strong pro-survival role in GBM, and is permanently activated in GBM, leading to increased cell proliferation, invasiveness, and angiogenesis. The aim of this work was to establish a GBM STAT3-knockout (STAT3-KO) cell line. The U87MG cell line and the CRISPR-Cas9 gene knockout method were used for this purpose. In a first step, we investigated the possibility of silencing STAT3 expression by siRNA and the suitability of this cell type to pinpoint the mechanism for CRISPR gene editing into the cell. In the second step, we transfected the cells using the STAT3 gene knockout kit (CRISPR) and tested the knockout success. In the established STAT3-KO GBM cell line, we confirmed STAT3-KO at the DNA, RNA and protein levels. We...
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Preparation of STAT3-knockout glioblastoma multiforme cell line using CRISPR-Cas9 editing
Papinčáková, Dominika ; Skálová, Lenka (advisor) ; Laštovičková, Lenka (referee)
Charles University Fakulty of Pharmacy in Hradec Králove Department of Biochemical Sciences Candidate: Bc. Dominika Papinčáková Supervisor: prof. RNDr. Lenka Skálová, Ph.D. Title of diploma thesis: Preparation of STAT3-knockout glioblastoma multiforme cell line using CRISPR-Cas9 editing Glioblastoma multiforme (GBM) is one of the most common primary brain tumours in adults and one of the most fatal cancers in humans A key oncogene in GBM development and progression is the signal transducer and activator of transcription 3 (STAT3). It is one of the mediators of tumorigenesis, plays a strong pro-survival role in GBM, and is permanently activated in GBM, leading to increased cell proliferation, invasiveness, and angiogenesis. The aim of this work was to establish a GBM STAT3-knockout (STAT3-KO) cell line. The U87MG cell line and the CRISPR-Cas9 gene knockout method were used for this purpose. In a first step, we investigated the possibility of silencing STAT3 expression by siRNA and the suitability of this cell type to pinpoint the mechanism for CRISPR gene editing into the cell. In the second step, we transfected the cells using the STAT3 gene knockout kit (CRISPR) and tested the knockout success. In the established STAT3-KO GBM cell line, we confirmed STAT3-KO at the DNA, RNA and protein levels. We...
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Determination of ovarian function parameters
Jará, Barbora ; Skálová, Lenka (advisor) ; Laštovičková, Lenka (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Thesis title: Determination of ovarian function parameters Supervisor: prof. RNDr. Lenka Skálová, Ph. D. Consultant: RNDr. Michala Vosátková, MBA Created by: Barbora Jará The levels of hormones that control of ovaria are important markers of development and health status of the female organism. Laboratory examination and correct interpretation of the results are important for proper diagnostic procedures. Among the hormones of ovarian function, the level of estradiol, progesterone, luteinizing and follicle-stimulating hormones is most often tested. Separation and immunochemical methods are used. For common clinical practice, immunochemical methods are particularly suitable due to their availability, relative accuracy, unpretentiousness, and speed of processing. The impossibility of standardizing individual methodological approaches represents the shortcoming of these methods. A series of examinations and interpretation of results can be performed using immunochemical methods only within one workplace. Keywords: estradiol, immunochemical methods, ECLIA, CMIA, RIA
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Influence of albendazole on the antioxidant system of alfalfa in real conditions
Lisoňková, Monika ; Szotáková, Barbora (advisor) ; Laštovičková, Lenka (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Monika Lisoňková Supervisor: prof. Ing. Barbora Szotáková, PhD. Title of diploma thesis: The effect of albendazole on the antioxidant system of alfalfa in real conditions Anthelmintics are drugs used to treat diseases caused by parasitics worms. In the Czech republic they are mainly used in veterinary medicine. Benzimidazole anthelmintic albendazole is used in cattle and sheep, during its biotransformation metabolites are formed, which are excreted together with the parent compound in faeces. Application of faeces from treated cattle to the field may affect non-target organisms, occurring in the field. An example of such organisms are agricultural crops, including alfalfa (Medicago sativa). Plants do not have the ability to eliminate the absorbed drugs and its metabolites, therefore they are deposited in plants and can affect the antioxidant system of plants. The aim of this study was to monitor the effect of albendazole on the antioxidant system of alfalfa (M. sativa) in real conditions. Plants were affected by faeces of sheep treated with albendazole. Subcellular fractions were prepared from harvested plants in the years 2019 and 2020. The concentrations of photosynthetic pigments...
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