636-0205-00L Lab Course: Mammalian Gene Circuits

Abstract	The students are trained in basic techniques in construction and characterization of synthetic gene circuits in mammalian cells. Experimental circuits are built with both the input and the output conjugated to fluorescent reporters, allowing characterization at the single cell level.
Learning objective	The objective of the course is to construct a genetic sensor for a molecular regulatory input such as microRNA or a transcription factor and characterize the input/output relationship of this sensor with the help of fluorescent reporters, fluorescent microscopy and fluorescent-activated cell sorting. The emphasis is on single-cell characterization.
Content	The course will take place over 4 weeks, with 2 days per week spent on lab work. The 4 weeks will be dedicated to the following activities Week 1: Introduction to the course; supervised construct design and detailed planning. Cloning of the constructs: part 1. Week 2: Cloning of the constructs, purification and characterization of DNA constructs Week 3: Cell culture transfection, microscopy and flow cytometry characterization Week 4: Data analysis and preparation of the final report; possibility to repeat failed experiments.
Lecture notes	Preparatory materials will be provided before the start of the course.
Literature	Will be provided before the course