Highly specific gene silencing by artificial microRNAs in the unicellular alga Chlamydomonas reinhardtii.
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.
Molnar
Attila
A
Department of Plant Sciences, University of Cambridge, Cambridge CB2 3EA, UK.
Bassett
Andrew
A
Thuenemann
Eva
E
Schwach
Frank
F
Karkare
Shantanu
S
Ossowski
Stephan
S
Weigel
Detlef
D
Baulcombe
David
D
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
D018076:DNA, Complementary; D035683:MicroRNAs; D015345:Oligonucleotide Probes; D012322:RNA Precursors; D018749:RNA, Plant