Genetically Modified Animals
Code: 100937 ECTS Credits: 6| Degree | Type | Year |
|---|---|---|
| 2500253 Biotechnology | OT | 4 |
Contact
- Name:
- Maria Fatima Bosch Tubert
- Email:
- fatima.bosch@uab.cat
Teachers
- Verónica Jimenez Cenzano
Teaching groups languages
You can view this information at the end of this document.
Prerequisites
There are no prerequisites to attend this course. However, to facilitate the student’s understanding of the subject matter and the achievement of the learning goals proposed, it is advisable that the student has previous knowledge on Cellular Biology, Genetics, Molecular Biology and Recombinant DNA technology.
It is also advisable that the students have basic knowledge of English, so that they can use the information sources of the field, which are mostly in this language.
Objectives and Contextualisation
The objective of the subject “Transgenic animals” is to provide the students with up-to-date knowledge in transgenesis and related technologies. Thus, the content of the subject will cover the following topics: Description and classification of transgenic animal models; Study of the different methodologies employed to obtain transgenic animal models of different species, and technologies that allow the overexpression of genes or the blockage or modification of endogenous genes, either ubiquitously or in a tissue-specific and/or inducible manner; Establishment and management of transgenic animal colonies; Cryopreservation of embryos and sperm, IVF, Health rederivation, Ethical aspects related to the generation and utilization of transgenic animals; Legislation on the use of laboratory animals; Application of animal transgenesis to the fields of biomedicine, biotechnology and livestock breeding.
Learning Outcomes
- CM35 (Competence) Evaluate the different methodologies useful for obtaining disease models.
- CM35 (Competence) Evaluate the different methodologies useful for obtaining disease models.
- CM37 (Competence) Apply the basic principles that regulate the interaction of drugs with organisms.
- KM38 (Knowledge) Detail the molecular bases of diseases and their various mechanisms.
- SM35 (Skill) Evaluate different molecular models or organisms for disease research.
Content
Knowledge on the following topics will be imparted during the theoretical classes:
TOPIC 1
Introduction to the technologies used for animal genetic engineering. Transgenic animals: definition and classification. Advantages of the mouse as an animal model in biomedicine.
TOPIC 2
Generation of transgenic animals by transgene addition. Preparation of DNA constructs or transgenes. Collection of embryos. Microinjection of DNA into the pronucleus of 1-cell embryos. Transfer of engineered embryos to receptor females. Genotyping of genetically engineered animals. Integration and vertical transmission of the transgene. Mosaic animals. Transgene expression and phenotype.
TOPIC 3
Design and production of chimeric genes/transgenes: promoters, inducible systems, insulators, enhancers. Analysis of transgene expression in vitro: technologies for the introduction of exogenous DNA to cultured cells. Transient and stable transfections. BACs and YACs.
TOPIC 4
Generation of transgenic livestock. Introduction of new traits of interest for livestock breeding. Biotechnological applications. Production of proteins with pharmaceutical interest in the mammary gland. Transgenic animals for xenotransplantation.
TOPIC 5
Generation of transgenic animals using viral vectors (lentivirus). Generation of transgenic animals from sperm. Transposons.
TOPIC 6
Targeted mutagenesis in animals through Embryonic Stem cells (ES cells): definition of ES cells, properties, obtainment and culture. Reprogramming and Induced Pluripotent Stem cells (iPS cells).
TOPIC 7
Generation of Knockout / Knockin mice by Gene targeting in ES cells. Design of recombination vectors. Homologous recombination. Selection of recombined ES clones.
TOPIC 8
Generation of mouse chimeras by injection of recombinant ES cells in blastocytes, injection/ aggregation of 8-cell embryos and tetraploid embryos. Homozygous and heterozygous Knockout / Knockin animals. Applications.
TOPIC 9
Conditional Knockout / Knockin animals: Recombinases systems (Cre-LoxP, FLP-Frt). Tissue-specific Knockout / Knockin animals. Inducible Knockout / Knockin animals; inducible systems, transcriptional and post-transcriptional control. Advantages and limitations. Applications.
TOPIC 10
Gene Trap for random mutagenesis. Technology and vectors for Gene Trap. Applications.
TOPIC 11
Knockout/in animal generation by Genome Edition. Zing Finger Nucleasas (ZFN), TALENs and CRISPR/Cas system. Advantages and limitations. Applications.
TOPIC 12
Generation of clonal animals: Nuclear transfer. Reprogramming. Applications. Advantages for the obtainment of transgenic livestock. Therapeutic cloning.
TOPIC 13
Establishment and maintenance of genetically modified mouse and rat colonies. Nomenclature. Phenotype: alterations arising due to transgenesis technology, environmental factors or genetic background.
TOPIC 14
Technologies to support the establishment and the management of colonies of genetically modified animals: Cryopreservation of embryos and sperm. In vitro fertilization (IVF). Health rederivation. Ovary transfer.
TOPIC 15
Current legislation on animal genetic engineering and use of laboratory animals. Ethical aspects. Ethics committees on animal experimentation.
TOPIC 16
Large International consortia on mouse mutagenesis. Large-scale phenotyping centres: “Mouse Clinics”.
TOPIC 17
Obtainment of transgenic fish. Applications in Biotechnology.
TOPIC 18
General Applications of Transgenic Animals.
TOPIC 19
Gene drive and control populations.
TOPIC 20
Applications of transgenic animals for the study of diseases (I): Diabetes mellitus and Obesity.
TOPIC 21
Applications of transgenic animals for the study of diseases (III): Hereditary metabolic diseases models.
TOPIC 22
Applications of transgenic animals for the study of diseases (IV): Neurosciences studies; Alzheimer and Parkinson diseases. Optogenetics.
The laboratory practice classes will cover the design of different types of transgenic animals and Knockout / Knockin mutants, the establishment and maintenance of colonies of transgenic mice and the genotypic analysis of the genetically engineered animals. Students will also carry out several techniques as part of the phenotypic analysis of genetically engineered mice. Using a transgenic mouse model, an in vivo phenotyping study will be performed.
Content of the laboratory practice classes:
- Generation of transgenic and Knockout / Knockin animals. Videos.
- Design of transgenes, gene targeting recombination vectors and components of the CRISPR/Cas9 system.
- Handling and in vitro culture of pre-implantational embryos.
- Genotype analysis. Establishment of colonies of transgenic animal and Knockout / Knockin mutants.
- Phenotype analysis. Histopathology, necropsy and in vivo studies.
Activities and Methodology
| Title | Hours | ECTS | Learning Outcomes |
|---|---|---|---|
| Type: Directed | |||
| Oral presentations | 8 | 0.32 | |
| Practical lessons | 12 | 0.48 | |
| Theorical lessons | 35 | 1.4 | |
| Type: Supervised | |||
| Tutorials | 5 | 0.2 | |
| Type: Autonomous | |||
| Individual study time | 74 | 2.96 | |
| Oral presentations | 10 | 0.4 |
The subject “Transgenic Animals” consists of theory and laboratory classes, and tutored oral presentations of relevant literature. The formative activities of the subject are complementary.
Theoretical classes
The contents of the theoretical classes will be imparted by a Professor in a series of master classes supported by audio-visual material. The slides used by each professor in each class will be available to the students through the subject’sCampus Virtual/Moodle. These master classes will constitute the main form of transfer of theoretical contents. Students are advised to periodically consult the books and links suggested in the Bibliography section of this document and at the Campus Virtual/Moodle to consolidate and clarify, if necessary, the contents explained in class.
Laboratory practice classes
The laboratory practice classes have been designed to help students get familiarized with the methodologies used to produce transgenic animals, establish animal colonies, genotype genetically engineered animals, and design and perform different phenotypic analyses in these animal models. We expect that, during these laboratory practice classes, students will be able to experience a “real world” situation in which they need to design an experiment, obtain a genetically engineered animal model and study in vivo their phenotype. We would like students to experience the excitement associated to the research that uses the technology of animal transgenesis.
The laboratory practice classes are composed of 3 sessions of 4 h each (from 3PM to 7PM), during which students will work in groups of 2-3 people under the supervision of an experienced professor. The date assigned to each laboratory practice group will be published in the subject’sCampus Virtual/Moodle with sufficient anticipation.
Attendance to laboratory practice classes is mandatory.
By the end of the laboratory practice classes, students will need to have answered a questionnaire. Both the laboratory practice guide and the questionnaire will be available through the Campus Virtual/Moodle. Students must bring their own lab coat, a waterproof marker and the Laboratory Practice Guide to each laboratory practice class.
Oral presentations of selected papers
Students will analyse and discuss in an oral presentation in front of the whole class a selected scientific publication on animal transgenesis, published in a recognized international scientific journal. To this end, students will pair with a fellow classmate. During the process of analysis of the paper’s content and preparation of the oral presentation, students will be tutored by researchers with experience in the field of animal transgenesis. Students will have 10 minutes for the oral presentation, equally divided amongst the members of the group, plus 5 minutes for questions (total of 15 minutes). The objective of this evaluating activity is that students get used -under the supervision of a tutor- to the process of searching, reading and understanding of scientific literature, and if necessary, develop a critical view on the figures, tables and results described in the publication. On the other hand, with this activity students will increase their knowledge of the current applications of the animal transgenesis technologies.
Tutoring
The oral presentations of selected papers will be tutored. In addition, upon request from the students, individualized tutoring will be available throughout the course. The objective of this sessions will be to help the student resolve doubts and reviewbasic concepts and to provide them with advice on sources of information and the best way to discuss scientific results in public.
UAB Surveys
15 minutes of one class will be allocated for the response of the UAB institutional survey.
Annotation: Within the schedule set by the centre or degree programme, 15 minutes of one class will be reserved for students to evaluate their lecturers and their courses or modules through questionnaires.
Assessment
Continous Assessment Activities
| Title | Weighting | Hours | ECTS | Learning Outcomes |
|---|---|---|---|---|
| Attendance to the oral presentations of research papers | 10% | 0 | 0 | CM35, KM38, SM35 |
| Examination of Laboratory classes | 15% | 1 | 0.04 | CM35, CM37 |
| Final examination of theoretical classes | 50% | 3 | 0.12 | CM35, SM35 |
| Oral presentations of selected research papers | 15% | 1 | 0.04 | CM35, KM38, SM35 |
| Self-study exercise | 10% | 1 | 0.04 | CM35, SM35 |
To pass the course, students must achieve a final score of 5 points (over a total of 10 points) and must attend the laboratory practice classes. The evaluation activities are:
1.- Final examination of theoretical classes
Accounts for 50% of the final score (5 points out of 10). Assessment will consist of a written examination, under the format of a True or False test, on topics explained during the theoretical classes. A score greater than 2.5 in this examination is required to pass the course.
There will be a Second Chance/Recovery Exam, under the same format as the original exam.
2.- Examination of Laboratory classes
Accounts for 15% of the final score (1.5 points out of 10). Assessment will consist of a written examination, under the format of a True or False test, on topics explained during the practical classes. It will be held the same day of the theoretical classes examination.
Attendance to practical sessions (or field trips) is mandatory. Students missing more than 20% of programmed sessions will be graded as "No Avaluable.
3.- Self-study exercise
Accounts for 10% of the final score (1 point out of 10). Assessment will consist of an exercise that the student will have to develop on their own. Details will be posted online in the "Campus Virtual" at the end of April.
4.- Oral presentations of selected research papers
Accounts for 15% of the final score (1.5 points out of 10). Students will be evaluated individually, both on their performance during the oral presentation of the selected paperand on the audio-visual material that they prepared to support their group presentation.
5.- Attendance to the oral presentations of research papers
Accounts up to 10% of the final score (1 point out of 10). Both attendance and participation in the scientific discussions of the sessions will be evaluated, following the scale:
Attendance 90-100% = 1 point
Attendance 80-89% = 0,8 points
Attendance 70-79% = 0,7 points
Attendance 60-69% = 0,6 points
Attendance 50-59% = 0,5 points
Attendance 0-49% = 0 points
6.-The review of the theory and practical exams qualifications will be carried out in person on a day and time communicated via the virtual campus, some days after the publication of the grades.
Single evaluation: The theory and practical exam will take place on the same day. Attendance to practical classes is mandatory. For the oral presentation of research papers, in the event that the ENTIRE GROUP that makes the presentation is eligible for the single evaluation, the group may make the oral presentation on the same day of the theoretical and practical exam, once the exam is finished. Students who take single assessment and who cannot attend the oral presentations, can optionally obtain the point of attendance to the oral presentations of selected research papers by performing an analysis of an article or an activity related to the topic of the subject, the same day of the exam once the exam is finished.
Bibliography
Bibliografy:
- Transgenic animals. Generation and use. L.M. Houdebine. Harwood Academic Publishers 1997.
- Mouse Genetics and Transgenics. A practical approach. Edited by: I.J. Jackson and C.M. Abbott. Oxford University Press. 2000. (www.oup.co.uk/PAS)
- Gene Targeting. A practical approach. Edited by: A.L. Joyner. Oxford University Press. 2000. (www.oup.co.uk/PAS)
- Manipulating the Mouse Embryo. A laboratory manual. (3rd Edition) Edited by: Andras Nagy et al. Cold Spring Harbor Laboratory Press. 2003.
- Transgenesis Techniques. Principles and Protocols. Edited by: Alan R. Clarke. Humana Press. 2002. (2nd Edition).
- Gene Knock-out Protocols. Edited by: Martin J. Tymms and Ismail Kola. Humana Press. 2001.
- Embryonic Stem Cells. Methods and Protocols. Edited by: Kursad Turksen. Humana Press. 2002.
- Human Molecular Genetics 2. T. Strachan i A.P. Read. John Wiley & Sons, Inc., Publication. 1999.
- Advanced Protocols for Animal Transgenesis. An ISTT Manual. Shirley Pease & Tomas L. Saunders (Editors). Springer. 2011.
- Editando genes: recorta, pega y colorea. Las maravillosas herramientas CRISPR. Lluís Montoliu. Colección el Café Cajal. Next Door Publishers.
2019
Interesting webs:
http://www.transtechsociety.org/
Software
Not applicable
Language list
| Name | Group | Language | Semester | Turn |
|---|---|---|---|---|
| (PLAB) Practical laboratories | 441 | Catalan/Spanish | second semester | afternoon |
| (PLAB) Practical laboratories | 442 | Catalan/Spanish | second semester | afternoon |
| (PLAB) Practical laboratories | 443 | Catalan/Spanish | second semester | afternoon |
| (PLAB) Practical laboratories | 444 | Catalan/Spanish | second semester | morning-mixed |
| (SEM) Seminars | 441 | Catalan/Spanish | second semester | morning-mixed |
| (TE) Theory | 44 | Catalan/Spanish | second semester | morning-mixed |