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This course will provide participants with hands-on tutorials and the theory behind gene transfer into fish embryo, as well as gene knock-in/-out using CRISPR/Cas. Microinjection into zebrafish and medaka embryos will be covered, as well as subsequent screening to identify positive founder fish.
This course is aimed at research technicians and scientists interested in the generation of genetic modified fish models. Participants should have basic experience with molecular biology techniques.
- Microinjection of lineage tracers into one-cell stage embryos
- Co-injection of DNA and mRNA into one-cell stage embryos
- Microinjection of CRISPR/Cas constructs
- Targeted inactivation of pigment loci or genes of interest provided by applicants (medaka and zebrafish)
- Homology directed repair in fish
- Screening by imaging
- High throughput, high content screening for successful events
- Molecular validation of Crispr/Cas9 mediated indel formation
The aim of the course is to provide participants with hands-on tutorials and the theory behind microinjection into medaka and zebrafish. In addition, participants will learn to design CRISPR constructs.