RNAi and Plant Gene Function Analysis: Methods and Protocols

ISBN-13: 9781617791222 / Angielski / Twarda / 2011 / 244 str.

Hiroaki Kodama; Atsushi Komamine

Opis

With RNAi technology now essential to most plant science researchers, this book introduces the central topics, lists the necessary materials and reagents, offers reproducible laboratory protocols, and provides notes on troubleshooting and avoiding pitfalls.

Spis treści

1. Overview of Plant RNAi

            Yuichiro Watanabe

 

2. Caveat of RNAi in Plants: the Off-Target Effect

            Muthappa Senthil-Kumar and Kirankumar S. Mysore

 

3. Plant Gateway Vectors for RNAi as a Tool for Functional Genomic Studies

            Toshiya Muranaka

 

4. Heat-Inducible RNAi for Gene Functional Analysis in Plants

            Frédéric Masclaux and Jean-Philippe Galaud

 

5. Gene Function Analysis by Artificial MicroRNAs in Physcomitrella patens

            Basel Khraiwesh, Isam Fattash, M. Asif Arif, and Wolfgang Frank

 

6. Virus-Induced Gene Silencing in Ornamental Plants

            Cai-Zhong Jiang, Jen-Chih Chen, and Michael Reid

 

7. Local RNA Silencing Mediated by Agroinfiltration

            Jutta Maria Helm, Elena Dadami, and Kriton Kalantidis

 

8. Direct Transfer of Synthetic Double-Stranded RNA into Protoplasts of Arabidopsis thaliana

            Ha-il Jung, Zhiyang Zhai and Olena K. Vatamaniuk

 

9. Detection of Long and Short Double-Stranded RNAs

            Toshiyuki Fukuhara, Syunichi Urayama, Ryo Okada, Eri Kiyota, and Hiromitsu Moriyama

 

10. Quantitative Stem-Loop RT-PCR for Detection of MicroRNAs

            Erika Varkonyi-Gasic and Roger P. Hellens

 

11. Large Scale Sequencing of Plant Small RNAs

            William P. Donovan, Yuanji Zhang, and Miya D. Howell

 

12. Computational Prediction of Plant miRNA Targets

            Ying-Hsuan Sun, Shanfa Lu, Rui Shi, and Vincent L. Chiang

 

13. Bisulfite Sequencing for Cytosine-Methylation Analysis in Plants

            Nazmul Haque and Masamichi Nishiguchi

 

14. Using Nuclear Run-On Transcription Assays in RNAi Studies

            Basel Khraiwesh

 

15. Proteomic Analysis of RNA-Interference Induced Knockdown Plant

            Sang Yeol Lee and Kyun Oh Lee

 

16. Comparative Analysis of Phosphoprotein Expression using 2D-DIGE

            Tomoya Asano and Takumi Nishiuchi

Blurb

The use of RNAi technology is essential for most plant science researchers. As DNA sequence information increases, so the need for functional annotation of target genes also increases. Authoritative and accessible, RNAi and Plant Gene Function Analysis: Methods and Protocols seeks to serve as an ideal guide to scientists of all backgrounds looking to further their knowledge of this fundamental science. Written in the highly successful Methods in Molecular Biology™ series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.

 

In addition to the functional annotation of unknown genes, RNAi technology has been applied to the genetic engineering of important plant metabolites including starches, oils, and storage proteins. RNAi has been used to engineer plants resistant to plant viruses and also to nematodes and insects. RNAi and Plant Gene Function Analysis: Methods and Protocols will provide both professionals and novices of plant biotechnology and breeding research with essential tips for designing experiments meant to explore plant gene function.