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Ribozymes And Sirna Protocols
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Book Synopsis Ribozymes and siRNA protocols by : Mouldy Sioud
Download or read book Ribozymes and siRNA protocols written by Mouldy Sioud and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 623 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.
Book Synopsis Ribozymes and siRNA protocols by : Mouldy Sioud
Download or read book Ribozymes and siRNA protocols written by Mouldy Sioud and published by Humana Press. This book was released on 2004-03-05 with total page 624 pages. Available in PDF, EPUB and Kindle. Book excerpt: In this completely updated and expanded edition of a classic bench manual, hands-on experts take advantage of the latest advances in ribozyme, DNAzyme, 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 hairpain 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 ribozymes protocols for effective design, selection, and therapeutic applications; and the latest RNAi methods for sequencing-specific gene silencing in a wide variety of organisms. Comprehensive and up-to-date, Ribozymes and siRNA Protocols synthesizes for experienced and novice investigators alike the exciting advances in understanding nucleic acid enzymes and demonstrates how they may be used to analyze gene function and target validation, and to productively develop new therapeutics for human diseases.
Book Synopsis Target Discovery and Validation Reviews and Protocols by : Mouldy Sioud
Download or read book Target Discovery and Validation Reviews and Protocols written by Mouldy Sioud and published by Springer Science & Business Media. This book was released on 2008-02-05 with total page 359 pages. Available in PDF, EPUB and Kindle. Book excerpt: These volumes review the most current methods for drug target discovery and validation. They explore how recent improvement in understanding the molecular mechanisms of human pathology is impacting drug target discovery in the laboratory and in real therapeutics, specifically for cancers and autoimmune disorders. This book provides a thorough review of the most cutting-edge methods available for each step in drug target identification, validation, and clinical application.
Book Synopsis RNA Towards Medicine by : Volker A. Erdmann
Download or read book RNA Towards Medicine written by Volker A. Erdmann and published by Springer Science & Business Media. This book was released on 2006-02-04 with total page 463 pages. Available in PDF, EPUB and Kindle. Book excerpt: Developments over the past few years have revealed the remarkable versatility of RNA in any compartment of the cell, tasks that had been thought to be exclusively in the realm of proteins and even beyond. The chapters in this book written by leading investigators in the field provide insight into various promising avenues where RNA and nucleic acid derivatives including antisense RNAs, such as siRNA, miRNAs, amplification/selection (SELEX) generated aptamers as well as ribozymes are at the threshold of impacting medicine.
Download or read book Cytokine Protocols written by Marc De Ley and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 248 pages. Available in PDF, EPUB and Kindle. Book excerpt: A collection of biochemical, cellular, and molecular techniques for unraveling and quantifying the events occurring between the initial contact of a cytokine at the membrane receptor and the eventual activation of gene transcription. The techniques used include the generation of transfectants, the immunohistochemical detection of cytokines in tissue sections, and optimized staining for cytoplasmic detection. Highlights include RT-PCR of small amounts of mRNA, in situ hybridization, biosensor analysis, measurement of biological activities and standardization, immunohistochemical and single-cell detection, and receptor isolation, characterization, and crystallization. Enjoy a quick and smooth introduction to the key methods used in cytokine research Use readily reproducible techniques that ensure successful experimental results Employ antisense-RNA, RT-PCR of small amounts of mRNA, and in situ hybridization.
Book Synopsis Drosophila Cytogenetics Protocols by : Daryl S. Henderson
Download or read book Drosophila Cytogenetics Protocols written by Daryl S. Henderson and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 467 pages. Available in PDF, EPUB and Kindle. Book excerpt: Leading drosophilists describe in step-by-step detail all the essential techniques for studying Drosophila chromosomes and suggest new avenues for scientific exploration. The chapters emphasize specimen preparation (from dissection to mounting) and cover both polytene and mitotic/meiotic chromosomes in depth. Each fully tested and readily reproducible protocol offers a background introduction, equipment and reagent lists, and tips on troubleshooting and avoiding pitfalls. A cutting-edge FISH and immunolocalization technique will be important for discovering how DNA sequence influences higher-order chromosome architecture and ultimately gene expression.
Book Synopsis Cell Cycle Checkpoint Control Protocols by : Howard B. Lieberman
Download or read book Cell Cycle Checkpoint Control Protocols written by Howard B. Lieberman and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 366 pages. Available in PDF, EPUB and Kindle. Book excerpt: The field of cell cycle regulation is based on the observation that the life cycle of a cell progresses through several distinct phases, G1, M, S, and G2, occurring in a well-defined temporal order. Details of the mechanisms involved are rapidly emerging and appear extraordinarily complex. Furthermore, not only is the order of the phases important, but in normal eukaryotic cells one phase will not begin unless the prior phase is completed successfully. Che- point control mechanisms are essentially surveillance systems that monitor the events in each phase, and assure that the cell does not progress prematurely to the next phase. If conditions are such that the cell is not ready to progress—for example, because of incomplete DNA replication in S or DNA damage that may interfere with chromosome segregation in M—a transient delay in cell cycle progression will occur. Once the inducing event is properly handled— for example, DNA replication is no longer blocked or damaged DNA is repaired—cell cycle progression continues. Checkpoint controls have recently been the focus of intense study by investigators interested in mechanisms that regulate the cell cycle. Furthermore, the relationship between checkpoint c- trol and carcinogenesis has additionally enhanced interest in these cell cycle regulatory pathways. It is clear that cancer cells often lack these checkpoints and exhibit genomic instability as a result. Moreover, several tumor suppressor genes participate in checkpoint control, and alterations in these genes are as- ciated with genomic instability as well as the development of cancer.
Book Synopsis Protein Purification Protocols by : Paul Cutler
Download or read book Protein Purification Protocols written by Paul Cutler and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 474 pages. Available in PDF, EPUB and Kindle. Book excerpt: The first edition of Protein Purification Protocols (1996), edited by Professor Shawn Doonan, rapidly became very successful. Professor Doonan achieved his aims of p- ducing a list of protocols that were invaluable to newcomers in protein purification and of significant benefit to established practitioners. Each chapter was written by an ex- rienced expert in the field. In the intervening time, a number of advances have w- ranted a second edition. However, in attempting to encompass the recent developments in several areas, the intention has been to expand on the original format, retaining the concepts that made the initial edition so successful. This is reflected in the structure of this second edition. I am indebted to Professor Doonan for his involvement in this new edition and the continuity that this brings. Each chapter that appeared in the original volume has been reviewed and updated to reflect advances and bring the topic into the 21st century. In many cases, this reflects new applications or new matrices available from vendors. Many of these have increased the performance and/or scope of the given method. Several new chapters have been introduced, including chapters on all the currently used protein fractionation and ch- matographic techniques. They introduce the theory and background for each method, providing lists of the equipment and reagents required for their successful execution, as well as a detailed description of how each is performed.
Book Synopsis Cell Migration in Inflammation and Immunity by : Daniele D’Ambrosio
Download or read book Cell Migration in Inflammation and Immunity written by Daniele D’Ambrosio and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 283 pages. Available in PDF, EPUB and Kindle. Book excerpt: Chemokines and their receptors play a central role in the pathogenesis of numerous, perhaps all, acute and chronic inflammatory diseases. About 50 distinct chemokines produced by a variety cell types and tissues either c- stitutively or in response to inflammatory stimuli are involved in a plethora of biological processes. These small secreted proteins exert their exquisitely variegated functions upon binding to a family of seven-transmembrane spanning G-protein coupled receptors (GPCRs) composed of almost 20 distinct entities. The biological activities of chemokines range from the control of leukocyte trafficking in basal and inflammatory conditions to the regulation of hema- poiesis, angiogenesis, tissue architecture, and organogenesis. The basis for such diversified activities rests, on one hand, upon the ubiquitous nature of chemokine production and chemokine receptor expression. Virtually every cell type can produce chemokines and expresses a unique combination of chemokine receptors. On the other hand, chemokine receptors make use of a flexible and complex network of intracellular signaling machineries that can regulate a variety of cellular functions ranging from cell migration, growth, and differentiation to death. As knowledge of the size of chemokine and chemokine receptor families rapidly reaches completeness, much is still to be uncovered in terms of fu- tional architecture of the chemokine system. The disparity between the large number of chemokines and that smaller number of receptors is balanced by the promiscuity in ligand–receptor interactions, with multiple chemokines binding to the same receptor and several chemokines binding to more than one receptor.
Book Synopsis Mammalian Artificial Chromosomes by : Vittorio Sgaramella
Download or read book Mammalian Artificial Chromosomes written by Vittorio Sgaramella and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 277 pages. Available in PDF, EPUB and Kindle. Book excerpt: In 1996, we organized a workshop, inter alia, at the National Research Co- cil in Milan under the generous sponsorship of the European Science Foun- tion. On that occasion, a small group of investigators convened from many countries and presented early evidence of the possibility of assembling basic units of mammalian chromosomes into artificial constructs (or, indeed, red- ing the relevant components to more manageable dimensions and defined c- stitution). Progress in the following years has been slow but steady. Many scientists who took part in the workshop have since been engaged in active and prod- tive research. It goes to the credit of Humana Press to have realized the need for a book on artificial chromosomes that aims to provide better tools to all scientists committed to this field who are confronted with very difficult tech- cal problems. We have strived to cover in Mammalian Artificial Chromosomes: Methods and Protocols all relevant areas of artificial chromosome research, from basic genetics to daring attempts to build new tools for genetic therapy. We are of course grateful to the authors who have accepted the task of describing the technical steps and pitfalls that can be encountered in their research. Rarely has a very delicate methodology been presented with such meticulous care. We have been helped in this enterprise by the excellent librarian of the LITA Institute in Segrate, Italy, Ms. Claudia Piergigli, whom we thank warmly. Ms.
Book Synopsis Atomic Force Microscopy by : Pier Carlo Braga
Download or read book Atomic Force Microscopy written by Pier Carlo Braga and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 388 pages. Available in PDF, EPUB and Kindle. Book excerpt: The natural, biological, medical, and related sciences would not be what they are today without the microscope. After the introduction of the optical microscope, a second breakthrough in morphostructural surface analysis occurred in the 1940s with the development of the scanning electron microscope (SEM), which, instead of light (i. e. , photons) and glass lenses, uses electrons and electromagnetic lenses (magnetic coils). Optical and scanning (or transmission) electron microscopes are called “far-field microscopes” because of the long distance between the sample and the point at which the image is obtained in comparison with the wavelengths of the photons or electrons involved. In this case, the image is a diffraction pattern and its resolution is wavelength limited. In 1986, a completely new type of microscopy was proposed, which, without the use of lenses, photons, or electrons, directly explores the sample surface by means of mechanical scanning, thus opening up unexpected possibilities for the morphostructural and mechanical analysis of biological specimens. These new scanning probe microscopes are based on the concept of near-field microscopy, which overcomes the problem of the limited diffraction-related resolution inherent in conventional microscopes. Located in the immediate vicinity of the sample itself (usually within a few nanometers), the probe records the intensity, rather than the interference signal, thus significantly improving resolution. Since the most we- known microscopes of this type operate using atomic forces, they are frequently referred to as atomic force microscopes (AFMs).
Book Synopsis Chiral Separations by : Gerald Gübitz
Download or read book Chiral Separations written by Gerald Gübitz and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 427 pages. Available in PDF, EPUB and Kindle. Book excerpt: Prominent experts from around the world detail the chromatographic and electroseparation techniques they have developed for chiral separations on an analytical scale. Described in step-by-step detail to ensure successful experimental results, the procedures are presented as either general methods or as specific applications to substance classes and special compounds, with emphasis on high performance liquid chromatography and capillary electrophoresis techniques, but also including thin layer chromotographic, gas chromatographic, supercritical fluid chromatographic as well as recent electrochromotographic techniques.
Book Synopsis HPLC of Peptides and Proteins by : Marie-Isabel Aguilar
Download or read book HPLC of Peptides and Proteins written by Marie-Isabel Aguilar and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 411 pages. Available in PDF, EPUB and Kindle. Book excerpt: The introduction of high-performance liquid chromatography (HPLC) to the analysis of peptides and proteins some 25 years ago revolutionized the biological sciences by enabling the rapid and sensitive analysis of peptide and protein structure through the exquisite speed, sensitivity, and resolution that can be easily obtained. Today, HPLC in its various modes has become the pivotal technique in the characterization of peptides and proteins and currently plays a critical role in both our understanding of biological processes and in the development of peptide- and protein-based pharmaceuticals. The number of applications of HPLC in peptide and protein purification continues to expand at an extremely rapid rate. Solid-phase peptide synthesis and recombinant DNA techniques have allowed the production of large quantities of peptides and proteins that need to be highly purified. HPLC techniques are also used extensively in the isolation and characterization of novel proteins that will become increasingly important in the postgenomic age. The design of multidimensional purification schemes to achieve high levels of product purity further demonstrates the power of HPLC techniques not only in the characterization of cellular events, but also in the production of pepti- and protein-based therapeutics. HPLC continues to be at the heart of the analytical techniques with which scientists in both academia and in industry must arm themselves to be able to fully characterize the identity, purity, and potency of peptides and proteins.
Book Synopsis G Protein Signaling by : Alan V. Smrcka
Download or read book G Protein Signaling written by Alan V. Smrcka and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 246 pages. Available in PDF, EPUB and Kindle. Book excerpt: Alan V. Smrcka presents a collection of cutting-edge methods for investigating G protein signaling from a variety of perspectives ranging from in vitro biochemistry to whole animal studies. Among the readily reproducible techniques presented are those for the purification of G proteins and effectors enzymes, assays of these purified G proteins and effector enzymes, and for the study of G proteins interactions with effectors in intact cells. Additional methods are provided for assaying G protein coupled receptor structure, function, and localization, and for studying the physiological roles for endogenous G proteins.
Book Synopsis Antibody Engineering by : Benny K. C. Lo
Download or read book Antibody Engineering written by Benny K. C. Lo and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 555 pages. Available in PDF, EPUB and Kindle. Book excerpt: The exquisite binding specificity of antibodies has made them valuable tools from the laboratory to the clinic. Since the description of the murine hybridoma technology by Köhler and Milstein in 1975, a phenomenal number of mo- clonal antibodies have been generated against a diverse array of targets. Some of these have become indispensable reagents in biomedical research, while others were developed for novel therapeutic applications. The attractiveness of an- bodies in this regard is obvious—high target specificity, adaptability to a wide range of disease states, and the potential ability to direct the host’s immune s- tem for a therapeutic response. The initial excitement in finding Paul Ehrlich’s “magic bullet,” however, was met with widespread disappointment when it was demonstrated that murine antibodies frequently elicit the human anti-murine an- body (HAMA) response, thus rendering them ineffective and potentially unsafe in humans. Despite this setback, advances in recombinant DNA techniques over the last 15–20 years have empowered the engineering of recombinant antibodies with desired characteristics, including properties to avoid HAMA. The ability to p- duce bulk quantities of recombinant proteins from bacterial fermentation also fueled the design of numerous creative antibody constructs. To date, the United States Food and Drug Administration has approved more than 10 recombinant antibodies for human use, and hundreds more are in the development pipeline. The recent explosion in genomic and proteomic information appears ready to deliver many more disease targets amenable to antibody-based therapy.
Book Synopsis Biopolymer Methods in Tissue Engineering by : Anthony P. Hollander
Download or read book Biopolymer Methods in Tissue Engineering written by Anthony P. Hollander and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 265 pages. Available in PDF, EPUB and Kindle. Book excerpt: There is an urgent need to develop new approaches to treat conditions as- ciated with the aging global population. The surgeon’s approach to many of these problems could be described as having evolved through three stages: Removal: Traditionally, diseased or badly damaged tissues and structures might simply be removed. This was appropriate for limbs and non-essential organs, but could not be applied to structures that were critical to sustain life. An additional problem was the creation of disability or physical deformity that in turn could lead to further complications. Replacement: In an effort to treat wider clinical problems, or to overcome the limitations of amputation, surgeons turned to the use of implanted materials and medical devices that could replace the functions of biological structures. This field developed rapidly in the 1960s and 1970s, with heart valve and total joint replacement becoming common. The term “biomaterial” was used increasingly to describe the materials used in these operations, and the study of biomaterials became one of the first truly interdisciplinary research fields. Today, biomaterials are employed in many millions of clinical procedures each year and they have become the mainstay of a very successful industry.
Book Synopsis Genetic Recombination by : Alan S. Waldman
Download or read book Genetic Recombination written by Alan S. Waldman and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 259 pages. Available in PDF, EPUB and Kindle. Book excerpt: Genetic recombination, in the broadest sense, can be defined as any process in which DNA sequences interact and undergo a transfer of information, producing new “recombinant” sequences that contain information from each of the original molecules. All organisms have the ability to carry out recombination, and this striking universality speaks to the essential role recombination plays in a variety of biological processes fundamentally important to the maintenance of life. Such processes include DNA repair, regulation of gene expression, disease etiology, meiotic chromosome segregation, and evolution. One important aspect of recombination is that it typically occurs only between sequences that display a high degree of sequence identity. The stringent requirement for homology helps to ensure that, under normal circumstances, a cell is protected from deleterious rearrangements since a swap of genetic information between two nearly identical sequences is not expected to dramatically alter a genome. Recombination between dissimilar sequences, which does happen on occasion, may have such harmful consequences as chromosomal translocations, deletions, or inversions. For many organisms, it is also important that recombination rates are not too high lest the genome become destabilized. Curiously, certain organisms, such as the trypanosome parasite, actually use a high rate of recombination at a particular locus in order to switch antigen expression continually and evade the host immune system effectively.
