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

SemesterSpring Semester 2020
LecturersY. Benenson
Periodicityyearly recurring course
Language of instructionEnglish


AbstractThe 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.
ObjectiveThe 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.
ContentThe 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 notesPreparatory materials will be provided before the start of the course.
LiteratureWill be provided before the course