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Gene Silencing

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David Crotty (crotty{at}cshl.edu; 516-422-4007), Executive Editor, Cold Spring Harbor Protocols

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Wayne Manos (manos{at}cshl.edu; 516-422-4009), Director, Serials Marketing, CSHL Press

Cost-effective method for gene silencing is featured in Cold Spring Harbor Protocols

COLD SPRING HARBOR, N.Y. (Wed., Aug. 1, 2007) – Nearly a decade ago, now-Nobel laureates Craig Mello and Andrew Fire discovered that they could insert short RNA molecules into worms and shut down specific genes. Today, scientists routinely use this powerful method, termed RNA interference, to study the functions of specific genes in mammalian systems.

In order to conduct these experiments, scientists generally rely on chemical synthesis of RNA molecules, which can be quite costly. A freely accessible article from this month’s release of Cold Spring Harbor Protocols (www.cshprotocols.org) addresses this problem; it describes a cost-effective approach for generating silencing RNAs, called esiRNAs, to efficiently target virtually any gene in mammalian cells.

The protocol (www.cshprotocols.org/cgi/content/full/2007/16/pdb.prot4824) describes how to enzymatically generate RNA molecules in vitro, using the cloned gene of interest as a template. The RNA molecules are then randomly cleaved into short fragments, purified, and used in RNA interference experiments.

The procedure was developed by Dr. Frank Buchholz’s group at the Max Planck Institute of Molecular Cell Biology and Genetics (Germany), and can be used to generate large sets of esiRNA libraries to be applied to large-scale studies of gene function (www.mpi-cbg.de/esiRNA/).

Also highlighted in Cold Spring Harbor Protocols this month is an article that describes how to culture thymus cells from fetal mice (www.cshprotocols.org/cgi/content/full/2007/16/pdb.prot4808). The thymus is the organ where T-cells—a principal component of the immune system in vertebrates—proceed through a strictly coordinated maturation process before being released into the bloodstream. Fetal thymus organ culture is the only system available for studying the complete program of T-cell maturation in vitro, and the protocol will be useful to researchers interested in understanding the intricacies of T-cell maturation. It was authored by Drs. Graham Anderson and Eric J. Jenkinson from the MRC Centre for Immune Regulation at the University of Birmingham (U.K.) (www.mrcbcir.bham.ac.uk/research/t-celldevelopment.htm).

Other articles published today include methods for imaging neuronal activity in zebrafish, examining gene expression patterns in fruit flies and frogs, preparing DNA from mammals for genotyping, and identifying protein-protein interactions in virtually any species. For a complete list of articles in the August release of Cold Spring Harbor Protocols, please see www.cshprotocols.org/TOCs/toc8_07.dtl.

About Cold Spring Harbor Protocols:
Cold Spring Harbor Protocols (www.cshprotocols.org)is an online resource of methods used in a wide range of biology laboratories. It is structured to be highly interactive, with each protocol cross-linked to related methods, descriptive information panels, and illustrative material to maximize the total information available to investigators. Each protocol is clearly presented and designed for easy use at the bench—complete with reagents, equipment, and recipe lists. Life science researchers can access the entire collection via institutional site licenses, and can add their suggestions and comments to further refine the techniques.

About Cold Spring Harbor Laboratory Press:
Cold Spring Harbor Laboratory Press is an internationally renowned publisher of books, journals, and electronic media, located on Long Island, New York. Since 1933, it has furthered the advance and spread of scientific knowledge in all areas of genetics and molecular biology, including cancer biology, plant science, bioinformatics, and neurobiology. It is a division of Cold Spring Harbor Laboratory, an innovator in life science research and the education of scientists, students, and the public. For more information, visit www.cshlpress.com.

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