title: The application of KillerRed for acute protein inactivation in living cells.
abstract: Generating loss of protein function is a powerful investigatory tool particularly if carried out on a physiologically relevant timescale in a live-cell fluorescent imaging experiment. KillerRed mediated chromophore assisted light inactivation (CALI) uses genetic encoding for specificity and light for acute inactivation that can also be spatially restricted. This unit provides protocols for setting up and carrying out properly controlled KillerRed experiments during live-cell imaging of cultured cells.
authors.affiliation: Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, Pennsylvania.
authors.forename: Timothy S
authors.initials: TS
authors.lastname: Jarvela
authors.forename: Adam D
authors.initials: AD
authors.lastname: Linstedt
mesh_terms: D000818:Animals; D002460:Cell Line; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D011993:Recombinant Fusion Proteins; C508771:killer red protein, Anthomedusae; D049452:Green Fluorescent Proteins
keywords: CALI; KillerRed; ROS; acute inactivation; protein inactivation