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.
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.
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.
Performance assessment information (valid until the course unit is held again)