Vectors and methods for hairpin RNA and artificial microRNA-mediated gene silencing in plants

Eamens, Andrew L. & Waterhouse, Peter M. (2011) Vectors and methods for hairpin RNA and artificial microRNA-mediated gene silencing in plants. Methods in Molecular Biology (Clifton, N.J.), 701, pp. 179-197.

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Abstract

In plant cells, DICER-LIKE4 processes perfectly double-stranded RNA (dsRNA) into short interfering (si) RNAs, and DICER-LIKE1 generates micro (mi) RNAs from primary miRNA transcripts (pri-miRNA) that form fold-back structures of imperfectly dsRNA. Both si and miRNAs direct the endogenous endonuclease, ARGONAUTE1 to cleave complementary target single-stranded RNAs and either small RNA (sRNA)-directed pathway can be harnessed to silence genes in plants. A routine way of inducing and directing RNA silencing by siRNAs is to express self-complementary single-stranded hairpin RNA (hpRNA), in which the duplexed region has the same sequence as part of the target gene's mRNA. Artificial miRNA (amiRNA)-mediated silencing uses an endogenous pri-miRNA, in which the original miRNA/miRNA* sequence has been replaced with a sequence complementary to the new target gene. In this chapter, we describe the plasmid vector systems routinely used by our research group for the generation of either hpRNA-derived siRNAs or amiRNAs.

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10 citations in Scopus
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12 citations in Web of Science®

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ID Code: 65913
Item Type: Journal Article
Refereed: Yes
Additional URLs:
Keywords: double stranded RNA, microRNA, plant RNA, small interfering RNA, article, gene vector, genetics, methodology, molecular cloning, molecular genetics, nucleotide sequence, plant, polymerase chain reaction, RNA interference, Base Sequence, Cloning, Molecular, Genetic Vectors, MicroRNAs, Molecular Sequence Data, Plants, RNA, Double-Stranded, RNA, Plant, RNA, Small Interfering
DOI: 10.1007/978-1-61737-957-4_10
ISSN: 1940-6029
Subjects: Australian and New Zealand Standard Research Classification > BIOLOGICAL SCIENCES (060000) > GENETICS (060400)
Australian and New Zealand Standard Research Classification > BIOLOGICAL SCIENCES (060000) > PLANT BIOLOGY (060700)
Divisions: Current > Schools > School of Earth, Environmental & Biological Sciences
Current > QUT Faculties and Divisions > Science & Engineering Faculty
Copyright Owner: Copyright 2011 John Wiley & Sons, Inc.
Deposited On: 09 Jan 2014 02:22
Last Modified: 14 May 2014 06:38

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