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| Title | Generating genetically modified mice for immunological research |
|---|---|
| Course number | 2186 |
| Programme | Allergi, immunologi och inflammation (Aii) |
| Language | English |
| Credits | 1.5 |
| Date | 2017-09-25 -- 2017-09-29 | Responsible KI department | Institutionen för medicin, Solna |
| Specific entry requirements | |
| Purpose of the course | The purpose of this course is to give the students an in depth theoretical understanding of technologies for generating precise genetic modifications in mice. The course focuses on traditional gene targeting techniques (homologous recombination in embryonic stem cells) and on newer techniques based on RNA-guided nucleases (i.e. CRISPR/Cas9) for non-homologous end joining (NHEJ) and homology directed repair (HDR). Understanding these technologies will enable the students to design experiments to test hypotheses in vivo and generate new tools to ask complex immunological questions. These skills are becoming ever more important as science is getting more complex. |
| Intended learning outcomes | After taking the course the student should have an in depth knowledge of how to generate genetically modified mice. The students should be able to write a scientifically sound gene targeting project plan at the end of the course. The student should acquire enough practical and theoretical knowledge to allow them to independently generate genetically modified mice. Specifically, the student should know how to design and make DNA constructs for classical gene targeting and for NEHJ and HDR using CRISPR/Cas9. The students should know how to design genotyping using e.g. Southern blotting and PCR, and finally how to use the modified mice in experiments. Furthermore, after the course the students should know how to critically analyse experiments presented in the scientific literature and judge their scientific quality. |
| Contents of the course | 1) Gene targeting and transgenesis in general: -A short history of gene targeting, transgenesis and CRISPR/Cas9 in mice -When are these technologies suitable for immunological experiments? -Overview of the work process. 2) Designing and making constructs for classical gene targeting, and for CRISPR/Cas9-based NHEJ and HDR. -How to obtain the necessary information for designing a gene targeting construct -Different approaches to make DNA constructs for gene targeting and CRISPR/Cas9-based NHEJ and HDR. 3) Conditional gene targeting. (Cre-lox system.) 4) Common problems in gene targeting, transgenesis, and CRISPR/Cas9; and how to solve them. 5) How to use genetically modified mice in immunological research. |
| Teaching and learning activities | Lectures will be the main form of teaching during the course. Workshops and a take-home examination are also critical elements of the course. In groups of three, the students will design a gene targeting (or CRISPR/Cas9) project and present their research plan. |
| Compulsory elements | The workshops and the take-home examination are compulsory. |
| Examination | Formative assessment during active participation in the workshop and summative assessment of the quality of the take-home examination in line with the intended learning outcomes of the course. Students will have a new examination opportunity within two months after the course is finished. |
| Literature and other teaching material | Recommended literature: A mouse geneticist's practical guide to CRISPR applications. Genetics. 2015 Jan;199(1):1-15. PMID: 25271304 Transgenic Mouse Methods and Protocols Methods in Molecular Biology Volume 693 2011 Editors: Marten H. Hofker, Jan van Deursen ISBN: 978-1-60761-973-4 |
| Number of students | 12 - 16 |
| Selection of students | Selection will be based on 1) the relevance of the course syllabus for the applicant's doctoral project (according to written motivation), 2) date for registration as a doctoral student (priority given to earlier registration date) |
| More information | |
| Additional course leader | |
| Latest course evaluation | Course evaluation report |
| Course responsible |
Alexander Espinosa Institutionen för medicin, Solna Alexander.Espinosa@ki.se |
| Contact person | - |
