551-1554-00L Multigene Expression in Mammalian Cells
|Semester||Spring Semester 2021|
|Lecturers||P. Berger, G. Schertler|
|Periodicity||yearly recurring course|
|Language of instruction||English|
|Comment||Number of participants limited to 5.|
The enrolment is done by the D-BIOL study administration.
General safety regulations for all block courses:
-Whenever possible the distance rules have to be respected
-All students have to wear masks throughout the course. Please keep reserve masks ready. Surgical masks (IIR) or medical grade masks (FFP2) without a valve are permitted. Community masks (fabric masks) are not allowed.
-The installation and activation of the Swiss Covid-App is highly encouraged
-Any additional rules for individual courses have to be respected
-Students showing any COVID-19 symptoms are not allowed to enter ETH buildings and have to inform the course responsible
|Abstract||Genetic engineering of mammalian cells with multiple expression cassettes is an essential need in contemporary cell biology. It is useful for protein expression for structural studies, the reprogramming of somatic cells, or for the expression of several fluorescently-tagged sensors. In this course, we use MultiLabel (Kriz et al., Nat. Commun., 2010) to create multigene expression plasmids.|
|Objective||Students will learn to design and clone multigene expression constructs for mammalian cells. The functionality of the constructs will be tested by immunofluorescence microscopy or Western blotting.|
|Content||We will clone fluorescently-tagged markers for subcellular compartments, assemble them to a multigene expression construct and transfect them into mammalian cells. These markers of subcellular compartments will be used to study the trafficking of activated receptors (e.g. serotonin receptor). Pictures will be taken on our microscopes and then we will quantify colocalization.|