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Cost-effective method for gene silencing is featured in Cold Spring Harbor Protocols
August 02, 2007Nearly 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 (http://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 (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 (http://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.) (http://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 http://www.cshprotocols.org/TOCs/toc8_07.dtl
Cold Spring Harbor Laboratory
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 (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 (http://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.) (http://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 http://www.cshprotocols.org/TOCs/toc8_07.dtl
Cold Spring Harbor Laboratory
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Rnai: A Guide to Gene Silencing (Manual) by Gregory J. Hannon (Author), Gregory J. Hannon (Editor) The ability of double stranded RNA to inhibit the expression of genes with an appropriate sequence can be harnessed to silence target genes in vitro in a new and powerful way. This volume combines reliable RNAi protocols for a variety of species with discussion of strategies for the effective design of experiments using this important new technique that is changing the way experimental science is done. |
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Gene Silencing: New Research by Grace W. Redberry (Editor) Gene silencing is a general term describing epigenetic processes of gene regulation. The term gene silencing is generally used to describe the 'switching off' of a gene by a mechanism other than genetic mutation. That is, a gene which would be expressed (turned on) under normal circumstances, is switched off by machinery in the cell. Genes are regulated at either the transcriptional or post-transcriptional level. Transcriptional gene silencing is the result of histone modifications, creating an environment of heterochromatin around a gene that makes it inaccessible to transcriptional machinery (RNA polymerase, transcription factors, etc.). Post-transcriptional gene silencing is the result of mRNA of a particular gene being destroyed. The destruction of the mRNA prevents translation to... |
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siRNA and miRNA Gene Silencing: From Bench to Bedside (Methods in Molecular Biology) by Mouldy Sioud (Editor) RNA interference has become a key method in the suppression of gene expression and the development of therapeutic agents, yet there is still the problem of delivery, stability, and the danger of off-target effects such as the silencing of unwanted genes and activation of innate immunity. In siRNA and miRNA Gene Silencing: From Bench to Bedside, expert researchers explore the most recent advances in siRNA design, expression, delivery, in vivo imaging, and methods to minimize siRNA’s unwanted effects and promote successful use in patients. As part of the highly successful Methods in Molecular Biology™ series, the chapters focus on their respective subjects with easy-to-use, up-to-date information, including several step-by-step laboratory protocols on topics such as new delivery... |
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Therapeutic Oligonucleotides: Transcriptional and Translational Strategies for Silencing Gene Expression (Annals of the New York Academy of Sciences) by Yoon S. Cho-Chung (Editor), Alan M. Gewirtz (Editor), Cy A. Stein (Editor) The potential for the development of therapeutic oligonucleotides into clinical medicines and their use as basic research tools are explored in this volume, which is the proceedings of the 7th NIH Symposium on Therapeutic Oligonucleotides. The focus is on antisense, RNAi, triple-helix, gene repair, DNA chips, and CpG immune modulatory oligonucleotides. Specific chapters address designing better siRNAs, splice switching oligonucleotides, selective delivery of oligonucleotides, and medicinal drugs by receptor-mediated endocytosis, development of a function overriding siRNA silencing in mammalian cells, transcription factor decoys, and modified oligonucleotide hybridization and genetic insertion. NOTE: Annals volumes are available for sale as individual books or... |
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An introduction to RNA-mediated gene silencing.: An article from: Science Progress by Mark Geanacopoulos (Author) This digital document is an article from Science Progress, published by Thomson Gale on March 22, 2005. The length of the article is 6704 words. The page length shown above is based on a typical 300-word page. The article is delivered in HTML format and is available in your Amazon.com Digital Locker immediately after purchase. You can view it with any web browser. Citation Details Title: An introduction to RNA-mediated gene silencing. Author: Mark Geanacopoulos Publication: Science Progress (Magazine/Journal) Date: March 22, 2005 Publisher: Thomson Gale Volume: 88 Issue: 1 Page: 49(21) Distributed by Thomson... | |
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Plant Gene Silencing by M.A. Matzke (Editor), A.J.M. Matzke (Editor) This book is an up-to-date and comprehensive collection of reviews on various aspects of epigenetic gene silencing in plants. Research on this topic has undergone explosive growth during the past decade and has revealed novel features of gene regulation and plant defense responses that also apply to animals and fungi. Gene silencing is relevant for agricultural biotechnology because stable expression of transgenes is required for the successful commercialization of genetically engineered crops. The reviews have been written by distinguished authors who have made significant contributions to plant gene silencing research. This volume supersedes other books on gene silencing by focussing on plant systems, where many pioneering experiments have been performed, and by including the... |
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Gene Silencing by RNA Interference: Technology and Application by Muhammad Sohail (Editor) Gene Silencing by RNA Interference: Technology and Application provides background in the field and describes methods and protocols for gene silencing. It offers a general introduction to the subject of gene silencing and RNA interference and focuses on technical details of the various methods of producing siRNAs and other RNAi tools. The final chapters of the book detail the application of RNAi to dissect gene function in a number of biological systems, including cell-free systems, cultured cells, and whole organisms. This book is a comprehensive guide to gene silencing by RNA interference methods for beginners and experienced professionals in the field. |
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Silencing genes could prove to be golden.(genetic manipulation in control of crown-gall disease)(Brief Article): An article from: On The Plate by Consumer Alert (Publisher) This digital document is an article from On The Plate, published by Consumer Alert on October 27, 2001. The length of the article is 347 words. The page length shown above is based on a typical 300-word page. The article is delivered in HTML format and is available in your Amazon.com Digital Locker immediately after purchase. You can view it with any web browser. Citation Details Title: Silencing genes could prove to be golden.(genetic manipulation in control of crown-gall disease)(Brief Article) Publication: On The Plate (Newsletter) Date: October 27, 2001 Publisher: Consumer Alert Page: NA Article Type: Brief Article Distributed by Thomson... | |
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Proceedings of the National Academy of Sciences of the United States of America November 10, 1998, Volume 95/ Number 23: Plant Gene Silencing Regularized; Bone Homeostasis by NAS (Author) | |
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Proceedings of the National Academy of Sciences of the United States of America July 21, 1998: Evidence for Gene Silencing; the Proton Collecting Function of the Inner Surface of Cytochrome C by NAS (Author) |
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