title: Highly specific gene silencing by artificial microRNAs in the unicellular alga Chlamydomonas reinhardtii.
abstract: MicroRNAs (miRNAs) are small RNAs, 21 to 22 nucleotides long, with important regulatory roles. They are processed from longer RNA molecules with imperfectly matched foldback regions and they function in modulating the stability and translation of mRNA. Recently, we and others have demonstrated that the unicellular alga Chlamydomonas reinhardtii, like diverse multicellular organisms, contains miRNAs. These RNAs resemble the miRNAs of land plants in that they direct site-specific cleavage of target mRNA with miRNA-complementary motifs and, presumably, act as regulatory molecules in growth and development. Utilizing these findings we have developed a novel artificial miRNA system based on ligation of DNA oligonucleotides that can be used for specific high-throughput gene silencing in green algae.
authors.affiliation: Department of Plant Sciences, University of Cambridge, Cambridge CB2 3EA, UK.
authors.forename: Attila
authors.initials: A
authors.lastname: Molnar
authors.forename: Andrew
authors.initials: A
authors.lastname: Bassett
authors.forename: Eva
authors.initials: E
authors.lastname: Thuenemann
authors.forename: Frank
authors.initials: F
authors.lastname: Schwach
authors.forename: Shantanu
authors.initials: S
authors.lastname: Karkare
authors.forename: Stephan
authors.initials: S
authors.lastname: Ossowski
authors.forename: Detlef
authors.initials: D
authors.lastname: Weigel
authors.forename: David
authors.initials: D
authors.lastname: Baulcombe
mesh_terms: D016825:Chlamydomonas reinhardtii / Q000235:genetics* / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D055785:Gene Knockdown Techniques / Q000379:methods*; D020868:Gene Silencing; D017343:Genes, Plant; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D059372:Nucleotide Motifs; D015345:Oligonucleotide Probes / Q000235:genetics / Q000378:metabolism; D059367:RNA Cleavage; D012322:RNA Precursors / Q000235:genetics / Q000378:metabolism; D020871:RNA Stability; D018749:RNA, Plant / Q000235:genetics / Q000378:metabolism*; D012680:Sensitivity and Specificity; D014170:Transformation, Genetic
chemical_list: D018076:DNA, Complementary; D035683:MicroRNAs; D015345:Oligonucleotide Probes; D012322:RNA Precursors; D018749:RNA, Plant