Vectors and Methods for Hairpin RNA and Artificial microRNA-Mediated Gene Silencing in Plants

Andrew Eamens; Peter M. Waterhouse

doi:10.1007/978-1-61737-957-4_10

Back

Vectors and Methods for Hairpin RNA and Artificial microRNA-Mediated Gene Silencing in Plants

Book chapter

Peer reviewed

Vectors and Methods for Hairpin RNA and Artificial microRNA-Mediated Gene Silencing in Plants

Andrew Eamens and Peter M. Waterhouse

Plant Chromosome Engineering, pp.179-197

Methods in Molecular Biology, 701, Humana Press

2011

DOI: https://doi.org/10.1007/978-1-61737-957-4_10

PMID: 21181531

Files and links (1)

url

https://doi.org/10.1007/978-1-61737-957-4_10View

Published Version

Abstract

amiRNA

dsRNA

hpRNA

miRNA

RNA silencing

siRNA

Plant Biology

Small RNA Biology

Arabidopsis thaliana

Molecular Manipulation

Gene Expression Regulation

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.

Details

Title: Vectors and Methods for Hairpin RNA and Artificial microRNA-Mediated Gene Silencing in Plants
Authors: Andrew Eamens (Author) - University of Sydney
Peter M. Waterhouse (Author) - University of Sydney
Contributors: James A Birchler (Editor) - University of Missouri
Publication details: Plant Chromosome Engineering, pp.179-197
Series: Methods in Molecular Biology; 701
Publisher: Humana Press
DOI: 10.1007/978-1-61737-957-4_10; 10.1007/978-1-61737-957-4
ISSN: 1940-6029
ISBN: 9781617379574
PMID: 21181531
Organisation Unit: School of Health and Behavioural Sciences - Legacy; University of the Sunshine Coast, Queensland; School of Health - Biomedicine
Language: English
Record Identifier: 99656589802621
Output Type: Book chapter

Metrics

26 Record Views

19 Times Cited - Web of Science

InCites Highlights

These are selected metrics from InCites Benchmarking & Analytics tool, related to this output

Web Of Science research areas: Biochemical Research Methods; Biochemistry & Molecular Biology; Plant Sciences

UN Sustainable Development Goals (SDGs)

This output has contributed to the advancement of the following goals:

Source: InCites