title: Pumilio-based RNA in vivo imaging.
abstract: Subcellular, sequence-specific detection of RNA in vivo is a powerful tool to study the macromolecular transport that occurs through plasmodesmata. The RNA-binding domain of Pumilio proteins can be engineered to bind RNA sequences of choice and fused to fluorescent proteins for RNA imaging. This chapter describes the construction of a Pumilio-based imaging system to track the RNA of Tobacco mosaic virus in vivo, and practical aspects of RNA live-cell imaging.
authors.affiliation: Biomedical Sciences Research Complex, University of St Andrews, BMS Building, North Haugh, St Andrews, Fife, KY16 9ST, UK, jt58@st-andrews.ac.uk.
authors.forename: Jens
authors.initials: J
authors.lastname: Tilsner
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D015870:Gene Expression; D005822:Genetic Vectors; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods*; D057054:Molecular Imaging; D008969:Molecular Sequence Data; D018515:Plant Leaves / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D030821:Plants, Genetically Modified; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D031425:Plasmodesmata / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D015202:Protein Engineering; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D012367:RNA, Viral / Q000032:analysis* / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D014027:Tobacco Mosaic Virus / Q000235:genetics* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D008164:Luminescent Proteins; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; C434479:citrine protein, bacteria