Directed Gene Silencing with Artificial MicroRNAs
The characterization of gene function typically includes a detailed analysis of loss-of-function alleles. In model plants, such as Arabidopsis thaliana and rice, sequence-indexed insertion collections provide a large resource of potential null alleles that can often be easily accessed through convenient Web sites (e.g., http://signal.salk.edu). They are, however, not available for nonmodel species, require stacking for knockout of redundant homologs, and do not easily allow for partial or regulated loss of gene function, which is particularly useful when null alleles are lethal. Transgene approaches that employ directed gene silencing can substitute for null alleles and also enable refined studies of gene function, e.g., by tissue-specific and inducible gene-silencing.
This chapter describes the generation and application of artificial microRNAs (amiRNAs) as a gene silencing tool in a wide variety of different plant species.
Key wordsGene silencing miRNA Hairpin Loss-of-function Phenotypic complementation
- 12.Warthmann N, Chen H, Ossowski O, Weigel D, Hervé P. Highly Specific Gene Silencing by Artificial miRNAs in Rice. submitted.Google Scholar
- 24.Tao Zhao, Wei Wang, Xue Bai and Yijun Qi; Gene silencing by artificial microRNAs in Chlamydomonas; The plant Journal, 2009Google Scholar
- 25.Attila Molnar, Andrew Bassett, Eva Thuenemann, Frank Schwach, Shantanu Karkare, Stephan Ossowski, Detlef Weigel and David Baulcombe; Highly specific gene silencing by artificial microRNAs in the unicellular alga Chlamydomonas reinhardtii; The Plant Journal, 2009Google Scholar
- 26.Basel Khraiwesh, Stephan Ossowski, Detlef Weigel, Ralf Reski, and Wolfgang Frank; Specific gene silencing by artificial MicroRNAs in Physcomitrella patens: an alternative to targeted gene knockouts. The Plant Journal, 2008.Google Scholar
- 27.Brodersen P, Sakvarelidze-Achard L, Bruun-Rasmussen M, Dunoyer P, Yamamoto YY, Sieburth L, Voinnet O, Widespread translational inhibition by plant miRNAs and siRNAs., Science. 2008 May 30;320(5880):1185–90Google Scholar