Highly Specific Gene Silencing by Artificial MicroRNAs in Arabidopsis[W][OA]

By R. Schwab, Stephan Ossowski, Markus Riester, Norman Warthmann, Detlef Weigel. The Plant Cell Online, 2006

1,426 citations recorded by the source corpus.

Plant microRNAs (miRNAs) affect only a small number of targets with high sequence complementarity, while animal miRNAs usually have hundreds of targets with limited complementarity. We used artificial miRNAs (amiRNAs) to determine whether the narrow action spectrum of natural plant miRNAs reflects only intrinsic properties of the plant miRNA machinery or whether it is also due to past selection against natural miRNAs with broader specificity. amiRNAs were designed to target individual genes or groups of endogenous genes. Like natural miRNAs, they had varying numbers of target mismatches. Previously determined parameters of target selection for natural miRNAs could accurately predict direct targets of amiRNAs. The specificity of amiRNAs, as deduced from genome-wide expression profiling, was as high as that of natural plant miRNAs, supporting the notion that extensive base pairing with targets is required for plant miRNA function. amiRNAs make an effective tool for specific gene silencing in plants, especially when several related, but not identical, target genes need to be downregulated. We demonstrate that amiRNAs are also active when expressed under tissue-specific or inducible promoters, with limited nonautonomous effects. The design principles for amiRNAs have been generalized and integrated into a Web-based tool (http://wmd.weigelworld.org).

Subfields: Plant Science

View Paper: https://www.semanticscholar.org/paper/286d32fe1461ccf145b0f9183e939d8fc8b43679

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Citations in the corpus

miR156-regulated SPL transcription factors define an endogenous flowering pathway in Arabidopsis thaliana. (2009, neutral): “In a complementary set of experiments, we tested the requirement for FUL activity in leaves and at the shoot apex by reducing its expression with an artiﬁcial miRNA (amiR-FUL) (Schwab et al., 2006).”
Development of Genetic and Genomic Research Resources for Brachypodium distachyon, a New Model System for Grass Crop Research (2008, neutral): “…mutation of candidate genes by TILLING (McCallum et al., 2000), delete-a-gene (Li and Zhang, 2002), RNAi or artifi cial microRNAs (amiRNAs) (Schwab et al., 2006), development of a full-length cDNA collection for recombinationbased manipulation of ORFs into epitope tags, yeast two-hybrid…”
microRNAs as oncogenes and tumor suppressors. (2007, background): “This was confirmed to be applicable by a recent study (Schwab et al., 2006).”
Antagonistic regulation of PIN phosphorylation by PP2A and PINOID directs auxin flux. (2007, background): “…seedlings were not recovered among progeny of pp2aa1 pp2aa2 pp2aa3/+ plants (n = 87), we constructed transgenic lines with estrogen (4-hydroxytamoxifen)-inducible overexpression of two different artificial microRNAs (amiRNAs) (Schwab et al., 2006) that simultaneously target all three PP2AA genes.”
Brassinosteroid signal transduction from cell surface receptor kinases to nuclear transcription factors (2009, methodology): “was transformed with an artificial microRNA construct targeting both BSL2 and BSL3 genes ( BSL 2,3-amiRNA), which was designed by the Web MicroRNA Designer 2 (ref”
Fungal Small RNAs Suppress Plant Immunity by Hijacking Host RNA Interference Pathways (2013, methodology): “Bc-sRNAs were cloned into the miRNA319a backbone vector (17) and transferred into the Gateway vector pEarley100 (without tag) for expression.”
The Balance between the MIR164A and CUC2 Genes Controls Leaf Margin Serration in Arabidopsis[W] (2006, background): “These findings suggest thatmiRNA processing is dependent on the hairpin structure of the pre-miRNA and extends to miRNAs expressed under control of their own promoter observations made previously for endogenous or artificial miRNAs expressed under the control of strong promoters, such as the 35S pro”
Elicitors, effectors, and R genes: the new paradigm and a lifetime supply of questions. (2007, background): “Virus-induced gene silencing (VIGS), artificial micro-RNAs (amiRNA), and mutation TILLING offer approaches to knock down gene expression in plants of economic interest, beyond Arabidopsis or Nicotiana benthamiana (34, 71, 142, 151), but the candidate receptor gene must first be identified.”
Gene silencing in plants using artificial microRNAs and other small RNAs. (2008, methodology): “Visible changes in amiRNA expression are readily detectable within 3 days with an ethanol-inducible system, and persist for several days after removal of the silencing trigger (but not on tissue that was initiated after the inductive pulse) (Schwab et al., 2006).”
Origin, biogenesis, and activity of plant microRNAs. (2009, neutral): “Likewise, overexpression of endogenous or artificial miRNA phenocopies the effects of knockout mutations despite appreciable accumulation of target mRNAs (Bari et al., 2006; Schwab et al., 2006).”
Functional diversity of miRNA in plants (2007, positive): “Finally, a better understanding on the miRNA regulation mechanisms in plants will make it possible to design artificial miRNAs that may be used as efficient tools for controlling gene expression at will [54,55].”