| Code |
16859
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| Year |
1
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| Semester |
S2
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| ECTS Credits |
5
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| Workload |
TP(40H)
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| Scientific area |
Biomedicina/Medicina/Bioquímica
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Entry requirements |
Not-applicable
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Learning outcomes |
The advanced course "From gene to protein: a practical approach" will allow students to apply this knowledge in the future in carrying out tasks related to the PhD project. Thus, at the end of the UC, the student should be able to:
a) Identify the different control points of gene expression and characterize the molecular mechanisms involved, as well as their regulation;
b) Describe the basic principles underlying gene expression analysis techniques by real-time PCR and Western blot and understand their use in different experimental contexts;
c) Perform laboratory procedures for real-time PCR and Western blot techniques;
d) Analyze and interpret data that result from gene expression analysis experiments;
e) Develop an experimental plan associated with one of the gene expression analysis techniques;
f) Develop critical capacity, analysis of scientific results ability and interaction with colleagues and professors collaboratively.
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Syllabus |
The contents below will be applied in the realization of a mini-project that aims to use a spermatogonial cell line (GC-spg1) to evaluate the effect of siRNA-mediated knockdown of estrogen receptor on the expression of c-kit, a gene associated with cell proliferation.
1. Regulation of gene expression: promoters, enhancers/silencers, insulators, mediators, coregulators, mechanisms of alternative splicing, interference RNA, miRNAs, lncRNAs, chromatin remodeling, and CpG methylation.
2. Advanced tools in DNA editing to control gene expression: siRNAs, shRNAs, and CRISPR.
3. Sample preparation for nucleic acid and protein extraction.
4. Real-time PCR: fundamentals, detection methods, primer design, analysis of primer specificity and efficiency, normalization, absolute and relative quantification, result analysis.
5. Western blot: fundamentals, detection methods, antibody selection, normalization, relative quantification, and result analysis.
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Main Bibliography |
Livro:
- Molecular Cell Biology, Lodish et al., 9th edition, New York : Macmillan Learning, 2021.
Artigos:
- DOI: 10.3390/ijms17030291
- DOI: 10.1093/nar/29.9.e45
- DOI: 10.1016/j.ab.2020.113608
- DOI: 10.1002/bmb.21516
- DOI: 10.1007/s10529-006-9127-2
- DOI: 10.3390/cells9071675
- DOI: 10.1016/j.bdq.2015.01.005
- DOI: 10.1186/1471-2199-11-74
- DOI: 10.2144/btn-2019-0065
- DOI: 10.2144/05391RV01
- DOI: 10.1007/s13353-013-0173-x
- DOI: 10.1038/nprot.2006.83
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Teaching Methodologies and Assessment Criteria |
The teaching methodologies are student-centered and aim to instill active learning through guided self-learning and discussion/interaction with colleagues during theoretical-practical (TP) and laboratory (PL) classes. Learning is developed in thematic seminars, complemented, and discussed during TP and PL classes. Appropriate resources are provided for the acquisition of the objectives of this advanced course. The evaluation will allow to assess the knowledge correctly and the technical skills acquired in the various activities. Objectives a) and b) are evaluated through a theoretical test. Objective c) is evaluated by observing the execution of laboratory techniques. Objective d) is evaluated in the theoretical test and through the presentation and articles discussion. Objective e) is evaluated through the design of a practical experiment. The objective f) is evaluated along the activities carried out by the interaction with colleagues.
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Language |
Portuguese. Tutorial support is available in English.
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