title: Single-Molecule Imaging of RNA Splicing in Live Cells.
abstract: Expression of genetic information in eukaryotes involves a series of interconnected processes that ultimately determine the quality and amount of proteins in the cell. Many individual steps in gene expression are kinetically coupled, but tools are lacking to determine how temporal relationships between chemical reactions contribute to the output of the final gene product. Here, we describe a strategy that permits direct measurements of intron dynamics in single pre-mRNA molecules in live cells. This approach reveals that splicing can occur much faster than previously proposed and opens new avenues for studying how kinetic mechanisms impact on RNA biogenesis.
authors.affiliation: Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Lisboa, Portugal.
authors.forename: José
authors.initials: J
authors.lastname: Rino
authors.affiliation: Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Lisboa, Portugal.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Martin
authors.affiliation: Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Lisboa, Portugal.
authors.forename: Célia
authors.initials: C
authors.lastname: Carvalho
authors.affiliation: Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Lisboa, Portugal.
authors.forename: Ana C
authors.initials: AC
authors.lastname: de Jesus
authors.affiliation: Instituto de Medicina Molecular, Faculdade de Medicina, Universidade de Lisboa, Lisboa, Portugal. Electronic address: carmo.fonseca@medicina.ulisboa.pt.
authors.forename: Maria
authors.initials: M
authors.lastname: Carmo-Fonseca
mesh_terms: D003587:Cytomegalovirus / Q000737:chemistry / Q000235:genetics; D004254:DNA Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D007438:Introns; D007700:Kinetics; D017909:Levivirus / Q000737:chemistry / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D057054:Molecular Imaging / Q000295:instrumentation / Q000379:methods*; D011401:Promoter Regions, Genetic; D012322:RNA Precursors / Q000235:genetics / Q000378:metabolism*; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D055544:beta-Globins / Q000235:genetics / Q000378:metabolism; D000097573:Red Fluorescent Protein
chemical_list: D004254:DNA Nucleotidyltransferases; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D012322:RNA Precursors; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D055544:beta-Globins; D000097573:Red Fluorescent Protein; C045119:FLP recombinase
keywords: Fluorescence microscopy; Live-cell imaging; RNA; Single molecule; Splicing