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Transgenic Plants pp 293–315Cite as

Expression of Artificial MicroRNAs in Physcomitrella patens

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 847))

Abstract

MicroRNAs (miRNAs) are ∼21-nt-long small RNAs transcribed from endogenous MIR genes which form precursor RNAs with a characteristic hairpin structure. MiRNAs control the expression of cognate target genes by binding to reverse complementary sequences resulting in cleavage or translational inhibition of the target RNA. Artificial miRNAs (amiRNAs) can be generated by exchanging the miRNA/miRNA* sequence of endogenous MIR precursor genes, while maintaining the general pattern of matches and mismatches in the foldback. Thus, for functional gene analysis, amiRNAs can be designed to target any gene of interest. During the last decade, the moss Physcomitrella patens emerged as a model plant for functional gene analysis based on its unique ability to integrate DNA into the nuclear genome by homologous recombination which allows for the generation targeted gene knockout mutants. In addition to this, we developed a protocol to express amiRNAs in P. patens that has particular advantages over the generation of knockout mutants and might be used to speed up reverse genetics approaches in this model species.

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Acknowledgments

This work was supported by Landesstiftung Baden-Württemberg (P-LS-RNS/40 to W.F.) and the German Academic Exchange Service (DAAD; PhD fellowships to I.F. and M.A.A.).

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Authors and Affiliations

  1. Plant Biotechnology, Faculty of Biology, University of Freiburg, Freiburg, Germany

    Isam Fattash & Wolfgang Frank

  2. Department of Plant Systems Biology, Flanders Institute for Biotechnology (VIB), Flanders, Belgium

    Basel Khraiwesh

  3. Department of Plant Biotechnology and Genetics, Ghent University, Gent, Belgium

    Basel Khraiwesh

  4. Institute for Genetics, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany

    M. Asif Arif

Authors
  1. Isam Fattash
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  2. Basel Khraiwesh
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  3. M. Asif Arif
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  4. Wolfgang Frank
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Correspondence to Wolfgang Frank .

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Editors and Affiliations

  1. School of Biological Sciences, University of Reading, Reading, Berkshire, RG6 6AS, United Kingdom

    Jim M. Dunwell

  2. Dept. of Agricultural Botany, School of Biological Sciences, University of Reading, Reading, United Kingdom

    Andy C. Wetten

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Fattash, I., Khraiwesh, B., Arif, M.A., Frank, W. (2012). Expression of Artificial MicroRNAs in Physcomitrella patens . In: Dunwell, J., Wetten, A. (eds) Transgenic Plants. Methods in Molecular Biology, vol 847. Humana Press. https://doi.org/10.1007/978-1-61779-558-9_25

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  • DOI: https://doi.org/10.1007/978-1-61779-558-9_25

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