Molecular basis of human genetic diseases
Sylabus zajęć
Informacje podstawowe
|
Okres
Semestr 1
|
Forma zajęć / liczba godzin / forma zaliczenia
|
Liczba punktów ECTS
3.0
|
Cele kształcenia dla zajęć
Wymagania wstępne
Academic knowledge of the genetics, biochemistry and molecular biology of the cell.Efekty uczenia się dla zajęć
| Symbol EU dla zajęć/przedmiotu | Po zakończeniu zajęć i potwierdzeniu osiągnięcia EU student/ka: | Symbole EK dla kierunku studiów |
| Efekt_01 | Know general laboratory safety procedures, according to BHP rules | K_W16, K_W17, K_K05, K_K09 |
| Efekt_02 | List human genetic diseases, describe theirs causes, molecular basis and symptoms | K_W04, K_U08 |
| Efekt_03 | Maintain mammalian cells in culture: growth, collection and transfection | K_W05, K_W06, K_U05, K_K05, K_K07, K_K04 |
| Efekt_04 | Describe strategies of transfection and transduction of mammalian cells | K_W05, K_W06, K_W11, K_W13 |
| Efekt_05 | Describe strategies of transient and stable knockdown (silencing), knockout (CRISPR/CAS9) and overexpression of genes and proteins respectively | K_W05, K_W10, K_W06, K_W07, K_W11, K_W12, K_W13, K_U08 |
| Efekt_06 | Describe methods for protein-protein and protein-RNA interactions and co-localization in vivo | K_W05, K_W06, K_W12, K_W13 |
| Efekt_07 | Prepare samples for IF, use microscope to see results, analysis of results | K_W04, K_W05, K_W10, K_W12, K_K05, K_K07, K_U04, K_U05, K_K05, K_K07, K_K04 |
| Efekt_08 | Describe cell cycle progression, methods used for cell cycle synchronization, reasons of cell cycle arrest, methods of cell cycle analysis | K_W04, K_W12, K_U04, K_U05, K_U08 |
| Efekt_09 | Describe prokaryotic and eukaryotic ribosomes, translation, application of polysome profiling | K_W12, K_U04, K_U05, K_U08 |
| Efekt_10 | Know available sources of scientific publications and how to use them to get necessary information | K_K02, K_K01, K_U08, K_W08 |
| Efekt_11 | Know how to prepare short scientific report based on the knowlegde aciured during the course | K_W05, K_W08, K_W13, K_U01, K_U03, K_U04, K_U07, K_U08, K_U09, K_K02, K_K03 |
Treści programowe
| Lp. | Treści programowe dla zajęć/przedmiotu | Symbol EU dla zajęć/przedmiotu |
| 1. |
Safe work in the laboratory |
Efekt_01 |
| 2. |
Examples of human genetic diseases, molecular basis of pathogenesis |
Efekt_02, Efekt_11 |
| 3. |
Type of cell lines, general rules for mammalian cell culture |
Efekt_03, Efekt_11 |
| 4. |
Different strategies for transfection and transduction of mammalian cells |
Efekt_03, Efekt_04, Efekt_11 |
| 5. |
Different strategies of knockdown (silencing), knockout (CRISPR/CAS9) and overexpression of genes and proteins |
Efekt_05, Efekt_11 |
| 6. |
Passaging of cells, counting of cells, seeding cells for transfection, inducing of shRNA production by doxycycline |
Efekt_03, Efekt_04, Efekt_05 |
| 7. |
Transfection of cells with plasmid encoding wild type FUS, ALS-associated FUS mutant and plasmid encoding polyglycine-GFP protein using Viromer and Lipofectamine 3000 transfection reagent |
Efekt_03, Efekt_04 |
| 8. |
Immunofluorescence staining of transfected cell: washing, fixation, incubation with primary and secondary antibodies, staining of nuclei using DAPI |
Efekt_06, Efekt_07 |
| 9. |
Basis of confocal microscope; methods of protein-protein and RNA-protein interaction analysis in vivo using confocal microscope (IF, FISH, PLA, RNA-PLA), visualization of optimal results using pre-made slides |
Efekt_06, Efekt_07, Efekt_11 |
| 10. |
Using the confocal microscopy to analyze the mislocalization of FUS protein within the cell and observation of poliglycine protein aggregation in the cytoplasm. Discussing the molecular consequences of genetic diseases, such as ALS and FXTAS. |
Efekt_02, Efekt_06, Efekt_07, Efekt_11 |
| 11. |
Cell cycle analysis: release cells from G2/M arrest, collecting cells in 4 time points, staining of cells with propidium iodide. |
Efekt_08 |
| 12. |
Analysis of their DNA content using flow cytometer. |
Efekt_08 |
| 13. |
Testing and discussing the influence of FUS knockout, knockdown, overexpression and mutation on cell cycle progression. |
Efekt_08, Efekt_05, Efekt_02, Efekt_11 |
| 14. |
Polysome profiling: cytoplasmic extract preparation, continuous glycerol gradient preparation, separation of ribosomes, monosomes and polysomes on gradient by ultracentrifugation, fractionation using BioComp system. |
Efekt_09, Efekt_11 |
| 15. |
Testing and discussing the influence of FUS knockout, knockdown, overexpression and mutation on ribosome assambly |
Efekt_05, Efekt_09, Efekt_02 |
| 16. |
Learning how to use available sources of scientific publications and write short scientific report based on the knowlegde aciured during the course |
Efekt_11, Efekt_10 |
Informacje dodatkowe
| Metody i formy prowadzenia zajęć |
|---|
| Sposoby oceniania | Symbole EK dla modułu zajęć/przedmiotu | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| EK_1 | EK_2 | EK_3 | EK_4 | EK_5 | EK_6 | EK_7 | EK_8 | EK_9 | EK_10 | EK_11 | ||
| Kryteria oceniania wg skali stosowanej w UAM | |
|---|---|
| bardzo dobry (bdb; 5,0): | |
| dobry plus (+db; 4,5): | |
| dobry (db; 4,0): | |
| dostateczny plus (+dst; 3,5): | |
| dostateczny (dst; 3,0): | |
| niedostateczny (ndst; 2,0): | |
Literatura
Wydawnictwa książkowe
- 1. BROWN T.A.: Genomes, Gerland Science Publishing, New York, USA, 2007
2. ALLISON L.A.: Fundamental Molecular Biology, Blacwell Publishing, Malden, USA, 2007
3. STRYER L.: Biochemistry, Macmillan Publishers, Basingstoke, UK, 2017
4. WĘGLEŃSKI P.: Genetyka molekularna, Wydawnictwo Naukowe PWN, Warszawa, Polska, 2017
Artykuły w czasopismach
- 1. Lagier-Tourenne, C., Polymenidou, M., Cleveland, D.W. (2010): TDP-43 and FUS/TLS: emerging roles in RNA processing and neurodegeneration, Hum. Mol. Genet., 19 R46-R64
2. Sama, R.R.K., Ward, C.L., Bosco, D.A. (2015): Functions of FUS/TLS From DNA Repair to Stress Response: Implications for ALS, ASN Neuro, 6
3. Bosco, D.A., Lemay, N., Ko, H.K., Zhou, H., Burke, C., Kwiatkowski, T.J., Sapp, P., McKenna-Yasek, D., Brown, R.H., Hayward, L.J. (2010): Mutant FUS proteins that cause amyotrophic lateral sclerosis incorporate into stress granules, Hum. Mol. Genet., 19, 4160-4175
4. Vance, C., Scotter, E.L., Nishimura, A.L., Troakes, C., Mitchell, J.C., Kathe, C., Urwin, H., Manser, C., Miller, C.C., Hortobágyi, T., et al. (2013): ALS mutant FUS disrupts nuclear localization and sequesters wild-type FUS within cytoplasmic stress granules, Hum. Mol. Genet., 22, 2676-2688
5. Romeo, V., Griesbach, E., Schümperli, D. (2014): CstF64: Cell Cycle Regulation and Functional Role in 3= End Processing of Replication-Dependent Histone mRNAs, Mol. Cell Biol., 34, 4272-4284
6. Krichevsky, A.M., Kosik K.S.. (2001): Neuronal RNA granules: a link between RNA localization and stimulation-dependent translation, Neuron, 32, 683-696
7. Jackman, J, .O’Connor, P.M. (1998): Methods for synchronizing cells at specific stages of the cell cycle, Curr. Prot. in Cell Biol., 8.3.1-8.3.20
8. Tate, S., Ferrigno, P.K. (2005): Cell cycle: synchronization at various stages, Encyclopedia of Life Sciences,
9. Hagerman, P.J., Hagerman, R.J. (2015): Fragile X-associated tremor/ataxia syndrome, Ann N Y Acad Sci ., 1338: 58–70
Nakład pracy studenta i punkty ECTS
| Forma aktywności | Średnia liczba godzin* na zrealizowanie aktywności | |
|---|---|---|
| Godziny zajęć (wg planu studiów) z nauczycielem | ||
| Praca własna studenta: | ||
| Przygotowanie do zajęć | ||
| Czytanie wskazanej literatury | ||
| Przygotowanie pracy pisemnej, raportu, prezentacji, demonstracji, itp. | ||
| Przygotowanie projektu | ||
| Przygotowanie pracy semestralnej | ||
| Przygotowanie do egzaminu / zaliczenia | ||
| SUMA GODZIN | ||
| LICZBA PUNKTÓW ECTS DLA MODUŁU ZAJĘĆ/PRZEDMIOTU | ||
* godzina (lekcyjna) oznacza 45 minut