Humana Ribozymes and siRNA protocols: 252 (Methods in Molecular Biology, 252)

During the last few years, new discoveries in the RNA field have opened up a wealth of opportunities to specifically target mRNA for functional analysis and target validation. Contained in this volume of Methods in Molecular Biology are methods useful for the design and application of ribozymes, DNAzymes, and small interfering RNAs (siRNAs). In addition, a number of illustrations aiming to facilitate the und- standing and application of each method is included. Like any new field, ribozymes and siRNA research is rapidly evolving, and so too are the methods used to study, select, express, and control their structure and in vivo biological activities. Ribozymes and siRNA Protocols, Second Edition focuses on the latest technical advances in interfering with gene expression. It contains a short int- duction followed by general and specific protocols for using hammerhead ribozymes and derivates, DNAzymes, hairpin ribozymes, group I intron ribozymes, RNase P ribozymes, and siRNAs. Also in this volume are general methods for RNA structure analysis, delivery of oligonucleotides, and gene therapy protocols using ribozymes. Review "...a timely and important addition to the literature in this rapidly expanding field...The vast scope of the book will appeal to experienced researchers as a source for new avenues of study, and as a central reference for the latest technologies." -Biochemist "...will be very useful for both novices and experts as a comprehensive collection of ribozyme protocols..." -Doody's Health Sciences Book Review Journal "...will provide a valuable source of reference for many laboratories..." -Microbiology Today From the Back Cover In this completely updated and expanded edition of a classic bench manual, hands-on experts take advantage of the latest advances in ribozyme, DNAzyme, hammerhead ribozymes and derivatives, and RNA interference technologies to describe in detail the exciting and successful methods now available for gene inactivation in vitro and in vivo. Their optimized techniques employ hairpin ribozymes, DNAzymes, hammerhead ribozymes and derivatives, group I intron ribozymes, RNase P ribozymes, and siRNAs, as well as general methods for RNA structure analysis, delivery of oligonucleotides, and gene therapy. Also provided are novel methods for identifying accessible cellular mRNA sites; group I intron and RNAse P ribozyme protocols for effective design, selection, and therapeutic applications; and the latest RNAi methods for sequence-specific gene silencing in a wide variety of organisms. Additional techniques cover the analysis of ribozyme structures and conformational transitions using nucleotide analog interference mapping and fluorescence resonance energy transfer, the use of ribozymes in clinical and gene therapy, and the use of ribozymes and DNAzymes in rodent models of human disease. Each proven protocol includes a background introduction outlining the principle behind the technique, step-by-step instructions, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and up-to-date, Ribozymes and siRNA Protocols details for experienced and novice investigators alike the many exciting advances in our understanding of nucleic acid enzymes, as well as demonstrating how they may be used to analyze gene function and target validation, and to productively develop novel therapeutics for human diseases.