The use of artificial MicroRNA technology to control gene expression in Arabidopsis thaliana

Eamens, Andrew L., McHale, Marcus, & Waterhouse, Peter M. (2014) The use of artificial MicroRNA technology to control gene expression in Arabidopsis thaliana. In Sanchez-Serrano, Jose J & Salinas, Julio (Eds.) Arabidopsis Protocols. Springer Science, New York ; London, pp. 211-224.

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In plants, double-stranded RNA (dsRNA) is an effective trigger of RNA silencing, and several classes of endogenous small RNA (sRNA), processed from dsRNA substrates by DICER-like (DCL) endonucleases, are essential in controlling gene expression. One such sRNA class, the microRNAs (miRNAs) control the expression of closely related genes to regulate all aspects of plant development, including the determination of leaf shape, leaf polarity, flowering time, and floral identity. A single miRNA sRNA silencing signal is processed from a long precursor transcript of nonprotein-coding RNA, termed the primary miRNA (pri-miRNA). A region of the pri-miRNA is partially self-complementary allowing the transcript to fold back onto itself to form a stem-loop structure of imperfectly dsRNA. Artificial miRNA (amiRNA) technology uses endogenous pri-miRNAs, in which the miRNA and miRNA(passenger strand of the miRNA duplex) sequences have been replaced with corresponding amiRNA/ amiRNAsequences that direct highly efficient RNA silencing of the targeted gene. Here, we describe the rules for amiRNA design, as well as outline the PCR and bacterial cloning procedures involved in the construction of an amiRNA plant expression vector to control target gene expression in Arabidopsis thaliana. © 2014 Springer Science+Business Media New York.

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2 citations in Scopus
5 citations in Web of Science®
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ID Code: 65846
Item Type: Book Chapter
Additional URLs:
Keywords: amiRNA, Arabidopsis, miRNA, Plant expression vector, RNA silencing, Target gene expression, microRNA, Arabidopsis thaliana, article, cloning vector, expression vector, gene amplification, gene expression regulation, gene silencing, molecular cloning, nonhuman, plant development, plant genetics, polymerase chain reaction, priority journal, sequence analysis
DOI: 10.1007/978-1-62703-580-4_11
ISBN: 9781627035798
Subjects: Australian and New Zealand Standard Research Classification > BIOLOGICAL SCIENCES (060000) > BIOCHEMISTRY AND CELL BIOLOGY (060100)
Australian and New Zealand Standard Research Classification > BIOLOGICAL SCIENCES (060000) > GENETICS (060400)
Divisions: Current > Schools > School of Earth, Environmental & Biological Sciences
Current > QUT Faculties and Divisions > Science & Engineering Faculty
Copyright Owner: Copyright 2014 Springer Science
Deposited On: 09 Jan 2014 08:04
Last Modified: 17 Feb 2016 03:28

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