Graduate School of health and Medical Sciences

Fish as experimental animals - zebrafish CRISPR and in vivo imaging
Provider: Faculty of Health and Medical Sciences

Activity no.: 3177-24-00-00
	Enrollment deadline: 20/05/2024

Date and time	30.05.2024, at: 08:00 - 07.06.2024, at: 18:00
Regular seats	16
Course fee	16,800.00 kr.
Lecturers	Louise von Gersdorff Jørgensen
ECTS credits	5.00

Contact person	Kirsten Grønlund Andersen E-mail address: kga@sund.ku.dk

Enrolment Handling/Course Organiser	PhD administration E-mail address: phdkursus@sund.ku.dk

Aim and content This course is free of charge for PhD students at Danish universities (except Copenhagen Business School), and for PhD Students from NorDoc member faculties. All other participants must pay the course fee. Anyone can apply for the course, but if you are not a PhD student at a Danish university, you will be placed on the waiting list until enrollment deadline. This also applies to PhD students from NorDoc member faculties. After the enrollment deadline, available seats will be allocated to applicants on the waiting list. Learning objectives To obtain basic knowledge on the biology of fish according to species-specific learning outcomes for EU module 3.1 (Basic and appropriate biology – species specific (theory)) and 3.2 (Basic and appropriate biology – species specific (practical)). To get informed on the use of fish as experimental animals and handling procedures according to species-specific learning outcomes for EU module 7 (Minimally invasive procedures without anaesthesia – species specific (theory) and 8 (Minimally invasive procedures without anaesthesia – species specific (skills)). To understand why the zebrafish is a popular model organism To obtain practical experience on husbandry and breeding using zebrafish according to species-specific learning outcomes for EU module 4 (Animal care, health and management – species specific (theory). To obtain knowledge on recognition of pain, suffering and distress in zebrafish according to species-specific learning outcomes for EU module 5 (Recognition of pain, suffering and distress - species specific). To obtain practical experience in microinjection of eggs with CRISPR/Cas reagents To obtain practical experience in in vivo real-time imaging in zebrafish larvae A student who has met the objectives of the course will: 1. Understand the biology of fish 2. Understand why zebrafish is a popular model 3. Be able to handle, anaesthetize and euthanize fish 4. Be able to conduct microinjections and gene manipulations of zebrafish using the CRISPR/Cas technology 5. Be able to conduct in vivo real time imaging of immune cells in zebrafish larvae 6. If the student has passed the laboratory animal science EU function ABD course, this course will function as an add on on fish and the student will be qualified to hold a license for making experiments on fish according to EU guidelines. If the student does not have the ABD course it will not be accredited as an add on. Content The course will cover a basic understanding of fish biology. Gills, body shape, skin, other organs, and basic immunology will be introduced to the students. The participants will also be taught how to anesthetize and euthanize fish as well as how to confirm anesthesia and death. The students will learn by attending lectures and by handling fish. To understand why the zebrafish is a unique model, a lecture will be given on husbandry including a description of biological characteristics advantageous for a vertebrate model organism. During this lecture examples will be provided of experiments where the zebrafish is utilized as an ideal model. The participants will have practical exercises handling fish, breeding zebrafish, collecting, and handling eggs and embryos. The students will get a basic introduction to the CRISPR/Cas technology. Thereafter, it will be shown which guide RNAs have been ordered – and why these were chosen - to knock out a gene responsible for pigmentation. Without this gene the fish becomes albino. The students will receive an introduction to microinjection of CRISPR reagents into newly fertilized zebrafish eggs, which include videos of the actual procedure in our laboratory. Furthermore, before they do the experimental procedure themselves, an instructor will demonstrate the technique. Two days following the injection the students themselves can verify their success rate with the CRISPR knockout by conducting visual inspection of the embryos and identifying albinos. One of the unique and valuable research tools in zebrafish is the in vivo real-time imaging. The larvae are transparent in early life and thousands of transgenic animals exist, where different components in the fish are tagged with fluorescence. This could be immune cells, receptors, the heart, or the whole vascular system. Therefore, we are capable of looking inside the animal while it is alive and record processes such as the heartbeat, immune cell behavior, receptor activation and much more. The students will receive an introduction to immobilize larvae by embedding them in low melting point agarose gel and subsequently they will conduct real-time imaging using a fluorescence stereo microscope. In the first week of the course the students will receive videos with lectures and instructions and be part of mini seminars online and in week two of the course (Tuesday 12 PM to Friday 12 PM) 2 half and 2 full days will be spent in the laboratories receiving instructions, doing practical exercises and mini-seminars. The exam will consist of 80% participation, 10 MCQs and a short assay lasting 1 hour where the students are allowed to use the materials they have received for the course. Participants All students or other interested personnel who are interested in working with fish either in aquaculture or with zebrafish as a model organism for other animals or humans. Furthermore, this course will target students who are interested in the CRISPR/Cas technology and its application. Relevance to graduate programmes The course is relevant to PhD students from the following graduate programmes at the Graduate School of Health and Medical Sciences, UCPH: - All graduate programmes Language English Form Lectures, discussions, seminars, practical exercises, instructions, and demonstrations. Course director Louise von Gersdorff Jørgensen Associate Professor Faculty of Health and Medical Sciences Section of parasitology and aquatic pathobiology lvgj@sund.ku.dk Teachers Louise von Gersdorff Jørgensen Associate Professor Faculty of Health and Medical Sciences Section of parasitology and aquatic pathobiology Sara Gelskov Research assistant Faculty of Health and Medical Sciences Section of parasitology and aquatic pathobiology Heidi Mathiessen PhD student Faculty of Health and Medical Sciences Section of parasitology and aquatic pathobiology Michael Le Pepke Postdoc Faculty of Health and Medical Sciences Center for Evolutionary Hologenomics Section of Hologenomics Ditte Goldschmidt Jepsen Master student Faculty of Health and Medical Sciences Section of parasitology and aquatic pathobiology Barbara Nowak Professor Institute for Marine and Antarctic Studies University of Tasmania Australia Moonika Haahr Marana Research Scientist The Center for Aquaculture Technologies (CAT) Canada Yajiao Duan Postdoc Faculty of Health and Medical Sciences Section of parasitology and aquatic pathobiology Dates 30-31 May 2023 + 4th-7th June 2024 (50 hours in total) Course location Stigbøjlen 7 1870 Frederiksberg C Registration Please register before 20 Mayl 2024 Expected frequency Once per year Seats to PhD students from other Danish universities will be allocated on a first-come, first-served basis and according to the applicable rules. Applications from other participants will be considered after the last day of enrolment. Note: All applicants are asked to submit invoice details in case of no-show, late cancellation or obligation to pay the course fee (typically non-PhD students). If you are a PhD student, your participation in the course must be in agreement with your principal supervisor.

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