Cost-Effective Method For Gene Silencing Is Featured In Cold Spring Harbor Proto

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Cost-Effective Method For Gene Silencing Is Featured In Cold Spring

Harbor Protocols

http://www.medicalnewstoday.com/articles/78540.php

Nearly a decade ago, now-Nobel laureates Craig Mello and 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 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 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. 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 (http://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

(see article). 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 and J. on from the MRC Centre

for Immune Regulation at the University of Birmingham (U.K.)

(http://www.mrcbcir.bham.ac.uk/research/t-celldevelopment.htm).

Other articles published 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.