Syllabus
Task 1: RNA interference of the embryonic lethal gene in C. elegans.
- C. elegans morphology, principle of the RNA interference
- in vitro synthesis of dsRNA
- preparation of microinjection tools (capillary, pads for the C. elegans immobilization)
- familiarization with the micromanipulator, microinjection process
Task 2: Study of migration potential of stem cells
- what are stem cells?
- meet Xenopus!
- preparation of RFP positive Sertoli stem cells derived from the testes of X. tropicalis juvenile male for the microinjection
- preparation of media, capillary and microinjection tools
- microinjection of Sertoli stem cells into X. tropicalis tadpoles
- light sheet microscopy - observation of migration routes of microinjected cells in living tadpoles
- preparation of photodocumentation
The course is taught with the support of the project reg. number CZ.02.2.69/0.0/0.0/16_015/0002362
Annotation
The aim of this practical course is to familiarize students with microinjection and micromanipulation techniques on the model of round worm (C. elegans) and clawed frogs (Xenopus laevis and Xenopus tropicalis). Students will have also the oportunity to work with advanced microscopic systems such as light sheet and live cell immaging microscopy.