title: Single-Molecule Live-Cell Visualization of Pre-mRNA Splicing.
abstract: Microscopy protocols that allow live-cell imaging of molecules and subcellular components tagged with fluorescent conjugates are indispensable in modern biological research. A breakthrough was recently introduced by the development of genetically encoded fluorescent tags that combined with fluorescence-based microscopic approaches of increasingly higher spatial and temporal resolution made it possible to detect single protein and nucleic acid molecules inside living cells. Here, we describe an approach to visualize single nascent pre-mRNA molecules and to measure in real time the dynamics of intron synthesis and excision.
authors.affiliation: Faculdade de Medicina, Instituto de Medicina Molecular, Universidade de Lisboa, Av. Prof. Egas Moniz, 1649-028, Lisbon, Portugal.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Martin
authors.affiliation: Faculdade de Medicina, Instituto de Medicina Molecular, Universidade de Lisboa, Av. Prof. Egas Moniz, 1649-028, Lisbon, Portugal.
authors.forename: José
authors.initials: J
authors.lastname: Rino
authors.affiliation: Faculdade de Medicina, Instituto de Medicina Molecular, Universidade de Lisboa, Av. Prof. Egas Moniz, 1649-028, Lisbon, Portugal.
authors.forename: Ana C
authors.initials: AC
authors.lastname: de Jesus
authors.affiliation: Faculdade de Medicina, Instituto de Medicina Molecular, Universidade de Lisboa, Av. Prof. Egas Moniz, 1649-028, Lisbon, Portugal. carmo.fonseca@medicina.ulisboa.pt.
authors.forename: Maria
authors.initials: M
authors.lastname: Carmo-Fonseca
mesh_terms: D058948:Cell Tracking / Q000379:methods*; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods*; D012322:RNA Precursors / Q000235:genetics*; D012326:RNA Splicing / Q000235:genetics*
chemical_list: D012322:RNA Precursors; D049452:Green Fluorescent Proteins
keywords: Live-cell imaging; Pre-mRNA; Single molecule; Spinning disk confocal microscopy; Splicing