@inbook{93804a04d4364938b216aade43940284,
title = "Chapter 16 How to Define Targets for Small Guide RNAs in RNA Silencing. A Biochemical Approach",
abstract = "RNA silencing involves various forms of sequence-specific gene silencing triggered by small RNAs. In RNA silencing, Argonautes are crucial protein components that are directed to the target messenger RNAs (mRNAs) through their association with small RNAs by base pairing. Argonautes repress the expression of the target genes at posttranscriptional levels. Full complementarity between a small RNA and its target mRNA results in Argonaute-mediated cleavage ({"}slicing{"}) of the target mRNA. The D-D-H (asparagine-asparagine-histidine) triad that exists in the PIWI domain of Argonautes is the catalytic center for rendering their target cleavage ({"}slicer{"}) activity. This chapter describes in vitro target RNA cleavage assays using Aubergine in a complex form with PIWI-interacting RNAs. Aubergine is one of the Argonautes expressed primarily in fly germ lines and is immunopurified from fly testes using the specific antibody against it. The method discussed is useful for defining targets for the small RNAs that function in RNA silencing.",
author = "Siomi, {Mikiko C.} and Nishida, {Kazumichi M.} and Haruhiko Siomi",
note = "Funding Information: We thank Keita Miyoshi for comments and advice on the manuscript. This work was supported by grants to M. C. Siomi and H. Siomi from Ministry of Education, Culture, Sports, Science and Technology of Japan, and the New Energy and Industrial Technology Development Organization. M. C. Siomi is also supported by CREST from JST. Mikiko C. Siomi is Associate Professor of Global COE for Human Metabolomics Systems Biology by MEXT.",
year = "2008",
doi = "10.1016/S0076-6879(08)02416-6",
language = "English",
series = "Methods in Enzymology",
publisher = "Academic Press Inc.",
number = "C",
pages = "345--355",
booktitle = "Methods in Enzymology",
address = "United States",
edition = "C",
}