Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.
Subscribe

Sci. STKE, 27 August 2002
Vol. 2002, Issue 147, p. pl13
[DOI: 10.1126/scisignal.1472002pl13]

PROTOCOLS

Confirming Specificity of RNAi in Mammalian Cells

Patrice Lassus*, Joe Rodriguez*, and Yuri Lazebnik*

The Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724, USA.

Abstract: RNA interference (RNAi) is a process of sequence-specific gene silencing. Recent advances in the understanding of RNAi have provided practical tools to silence gene expression in mammalian cells, opening new possibilities for studying the functions of genes and proteins. It is important to ensure that an observed effect of RNAi is due to silencing of the intended target. Indeed, it is possible that an siRNA may silence more than one messenger RNA that is homologous in the region complementary to the siRNA. Considering that we know little about how RNAi works in mammalian cells, other artifacts may be yet to be recognized. Thus, we suggest approaches to rescue the effect of RNAi by ectopically expressing the protein of interest. These approaches involve introducing silent mutations into the complementary DNA of the protein and targeting RNAi to the untranslated regions of the gene.

*Corresponding authors. E-mails: lassus{at}cshl.edu (P.L.), rodrigue{at}cshl.org (J.R.), lazebnik{at}cshl.edu (Y.L.).

Citation: P. Lassus, J. Rodriguez, Y. Lazebnik, Confirming Specificity of RNAi in Mammalian Cells. Sci. STKE 2002, pl13 (2002).

Read the Full Text

THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
The TGF{beta} Receptor-interacting Protein km23-1/DYNLRB1 Plays an Adaptor Role in TGF{beta}1 Autoinduction via Its Association with Ras.
Q. Jin, W. Ding, and K. M. Mulder (2012)
J. Biol. Chem. 287, 26453-26463
   Abstract »    Full Text »    PDF »
RNA interference rescue by bacterial artificial chromosome transgenesis in mammalian tissue culture cells.
R. Kittler, L. Pelletier, C. Ma, I. Poser, S. Fischer, A. A. Hyman, and F. Buchholz (2005)
PNAS 102, 2396-2401
   Abstract »    Full Text »    PDF »

To Advertise     Find Products

Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882