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 BiologyTM 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 formulations and strategies with promising applications in vitro and in vivo, validated therapeutic target genes, and components of miRNA function, biogenesis, and interference with virus infection. Comprehensive and cutting-edge, siRNA and miRNA Gene Silencing: From Bench to Bedside offers an excellent collection of chapters to aid all those with an interest in RNAi, gene regulation, and new therapies.
"Small interfering RNA (siRNA) and microRNA (miRNA) are short double stranded RNA molecules that mediate posttranscriptional gene silencing via RNA interference by binding to or degrading complimentary target mRNA in the cytosol to inhibit translation. Though > 30 siRNA and miRNA candidates are in clinical trials for a variety of indications, none have yet to receive FDA-approval. Successful delivery is the critical barrier to therapeutic translation, and necessitates the need for multifunctional drug delivery systems (DDS). Barriers to delivery include nuclease-mediated degradation and phospholipid membrane impermeability. Furthermore, any DDS used must be cytocompatible and avoid off-target effects. The study herein provides a comprehensive characterization of a pH-responsive diblock copolymer nanoparticle (NP) siRNA DDS and its effects on mesenchymal stem cells (MSCs) for applications in regenerative medicine. Reversible addition-fragmentation chain transfer (RAFT) polymerization was used to synthesize a library of diblock copolymers to determine how polymer properties affect siRNA delivery. Results show this NP-siRNA delivery is effective in multiple therapeutic cell types in vitro, and that increasing NP core hydrophobicity provides greater cytocompatibility and gene silencing. This is achieved by increased pHresponsive membrane lytic activity allowing for better endosomal escape. Additionally, in vitro treatment conditions were identified that resulted in potent gene silencing with no acute cytotoxicity. However, NP-siRNA treatment resulted in a sustained reduction in hMSC metabolic activity, and RNAseq with enrichment analysis shows upregulation of anti-apoptotic gene expression and immune signaling pathways associated with siRNA immune stimulation. Colloidal stability is critical for effective NP DDS, and can be significantly altered in the presence of biological proteins. Therefore, an extensive characterization of this NP-siRNA DDS in the presence of serum reveals NP-siRNA aggregation, which significantly diminishes siRNA delivery in vitro. When delivered locally in vivo, cells take up NP-siRNA complexes in a non-aggregated state, emphasizing the importance of local delivery. Taken together this in-depth characterization of NP-mediated siRNA delivery establishes a solid foundation for the design of next generation NP-siRNA DDS for the delivery of siRNA/miRNA to MSCs."--Pages xiv-xv.
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 form an active gene product (in most cases, a protein). A common mechanism of post-transcriptional gene silencing is RNAi. Both transcriptional and post-transcriptional gene silencing are used to regulate endogenous genes. This book presents the latest research in this important field.
RNAi technology is used for large-scale screens that systematically shut down each gene in the cell, which can help identify the components necessary for a particular cellular process or an event such as cell division. Exploitation of the pathway is also a promising tool in biotechnology and medicine. Introducing new technology in the study of RNA
RNA technologies are the driving forces of modern medicine and biotechnology. They combine the fields of biochemistry, chemistry, molecular biology, cell biology, physics, nanotechnology and bioinformatics. The combination of these topics is set to revolutionize the medicine of tomorrow. After more than 15 years of extensive research in the field of RNA technologies, the first therapeutics are ready to reach the first patients. Thus we are witnessing the birth of a very exciting time in the development of molecular medicine, which will be based on the methods of RNA technologies. This volume is the first of a series. It covers various aspects of RNA interference and microRNAs, although antisense RNA applications, hammerhead ribozyme structure and function as well as non-coding RNAs are also discussed. The authors are internationally highly respected experts in the field of RNA technologies.
This special volume of The Enzymes is targeted towards researchers in biochemistry, molecular and cell biology, pharmacology, and cancer. This thematic volume discusses Eukaryotic RNases and their partners in RNA degradation and biogenesis. Contributions from leading authorities Informs and updates on all the latest developments in the field
Currently, intensive effort is being directed toward the identification of molecular targets that can provide approaches to the development of novel therapeutic strategies in cancer management. This book focuses on metastasis-associated genes, metastasis promoter and suppressor genes, which relate specifically to behavioral alterations of cancer cells in epithelial mesenchymal transition, cancer stem cell maintenance and propagation, and to the acquisition of invasive and metastasis faculty. The function of these genes has implications for cell cycle regulation and cell proliferation and so constitute an essential element in cancer growth and dissemination. The emphasis in this book is on how appropriate these genes are as molecular targets and how practicable are the constituents of their signal transduction systems as potential candidates and how accessible they are to targeted therapy. Written in a straightforward and clear style with background information supporting the new research, this book will be useful for students and researchers in cancer therapies. Identifies molecular targets and their accessibility for therapeutic intervention Provides information on biological features of tumor development and dissemination Background information provided for each topic
Commonly used by researchers to develop technologies for modifying and studying genetic process, RNA interference (RNAi) has many potential uses in medicine, biotechnology, and functional genomics. This book covers all essential aspects involved in the development of RNAi therapeutics, providing detailed guidance on the challenges and opportunities of bringing RNAi technologies from bench to clinic. It explores the design and mechanism of RNAi molecules, delivery strategies, and therapeutic applications in various diseases. Preclinical, regulatory, market, and intellectual aspects of RNAi technologies are also covered.
In the most recent years, each of the RNA silencing pathways of plants have appeared to generate ncRNAs with dedicated functions, specialized biological activities and specific functional scopes. RNA silencing plays a crucial role in coordinating the expression, stability, protection and inheritance of eukaryotic genomes. It compromises several mechanisms, that invariably depend on core small non coding RNAs and that achieve dedicated sequence-specific functions. RNA silencing has been recognized to carry critical developmental, stress-response and bodyguard functions be coordinating the expression, protection, stability and inheritance of virtually all eukaryotic genomes. Thus, the ncRNAs encompass a wide set of mechanisms that achieve specialized functions.
This book focuses on an emerging, central issue in molecular genetics and the development of eukaryotes: the control of gene expression by small species of RNA. As an exciting new field of endeavor, it is the first book by a single author to deal comprehensively with RNA silencing.The book provides the historical background of the field preceding the seminal work by Fire and associates in 1998 on the impact of small double-stranded RNA on the expression of nematode genes, which is considered the beginning of RNA silencing research. RNA silencing is described in a wide range of plants and animals including protozoa, simple metazoa, insects, non-mammalian vertebrates, and mammals. In each case the experimental results are provided with the accompanying background and with illustrations. There is also an appendix on the prospective use of RNA silencing in gene therapy, which is intended as a guide for investigators wishing to explore this possibility.
Leguminous crops have been found to contribute almost 27% of the world’s primary crop production. However, due to environmental fluctuations, legumes are often exposed to different environmental stresses, leading to problems with growth and development, and ultimately, decreased yield. This timely review explains the transcriptomics, proteomics, genomics, metabolomics, transgenomics, functional genomics and phenomics of a wide range of different leguminous crops under biotic and abiotic stresses, and their genetic and molecular responses. Amongst others the text describes the effect of nutrient deficiency, pesticides, salt, and temperature stress on legumes. Importantly, the book explores the physiobiochemical, molecular and omic approaches that are used to overcome biotic and abiotic constraints in legumes. It looks at the exogenous application of phytoprotectants; the role of nutrients in the alleviation of abiotic stress; and the microbial strategy for the improvement of legume production under hostile environments. Key features: demonstrates how to mitigate the negative effect of stress on leguminous crops, and how to improve the yield under stress the most up-to-date research in the field written by an international team of active researchers and practitioners across academia, industry and non-profit organisations. This volume is a valuable and much-needed resource for scientists, professionals and researchers working in plant science, breeding, food security, crop improvement and agriculture worldwide. In universities it will educate postgraduate and graduate students in plant science and agriculture; it will also benefit those in scientific institutions and in biotech and agribusiness companies, who deal with agronomy and environment.
Plant gene silencing is a crucially important phenomenon in gene expression and epigenetics. This book describes the way small RNA is produced and acts to silence genes, its likely origins in defence against viruses, and also its potential to improve plants. Plant gene silencing can be used to improve industrial traits, make plants more nutritious or more valuable to consumers, to remove allergens, and to improve resistance to weeds and pathogens.
Complete, up-to-date coverage of the broad area of nucleic acid chemistry and biology Assembling contributions from a collection of authors with expertise in all areas of nucleic acids, medicinal chemistry, and therapeutic applications, Medicinal Chemistry of Nucleic Acids presents a thorough overview of nucleic acid chemistry—a rapidly evolving and highly challenging discipline directly responsible for the development of antiviral and antitumor drugs. This reliable resource delves into a multitude of subject areas involving the study of nucleic acids—such as the new advances in genome sequencing, and the processes for creating RNA interference (RNAi) based drugs—to assist pharmaceutical researchers in removing roadblocks that hinder their ability to predict drug efficacy. Offering the latest cutting-edge science in this growing field, Medicinal Chemistry of Nucleic Acids includes: In-depth coverage of the development and application of modified nucleosides and nucleotides in medicinal chemistry A close look at a large range of current topics on nucleic acid chemistry and biology Essential information on the use of nucleic acid drugs to treat diseases like cancer A thorough exploration of siRNA for RNAi and the regulation of microRNA, non-coding RNA (ncRNA), a newly developing and exciting research area Thorough in its approach and promising in its message, Medicinal Chemistry of Nucleic Acids probes the new domains of pharmaceutical research—and exposes readers to a wealth of new drug discovery opportunities emerging in the dynamic field of nucleic acid chemistry.
In the last few years the major effect that RNAi has had in invertebrate systems is beginning to take hold in mammalian systems through both single gene knockdown experiments and genome-scale screens. In the next decade, there will no doubt be both notable successes and failures as we attempt to apply this genetic tool to various biological problems. Through the introduction of RNAi, mammalian systems have finally gained admittance to the pantheon of model genetic systems.
Animal genetics is a foundational discipline in the fields of animal science, animal breeding, and veterinary sciences. While genetics underpins the healthy development and breeding of all living organisms, this is especially true in domestic animals, specifically with respect to breeding for key traits. Molecular and Quantitative Animal Genetics is a new textbook that takes an innovative approach, looking at both quantitative and molecular breeding approaches. The bookprovides a comprehensive introduction to genetic principles and their applications in animal breeding. This text provides a useful overview for those new to the field of animal genetics and breeding, covering a diverse array of topics ranging from population and quantitative genetics to epigenetics and biotechnology. Molecular and Quantitative Animal Genetics will be an important and invaluable educational resource for undergraduate and graduate students and animal agriculture professionals. Divided into six sections pairing fundamental principles with useful applications, the book's comprehensive coverage will make it an ideal fit for students studying animal breeding and genetics at any level.
This methods manual provides an introduction to RNA interference, the theory behind its many applications, and specific protocols for RNAi, in organisms from plants and C.elegans to Drosophila and mammals. There are also chapters covering small hairpin RNAs and viral-induced gene silencing.
RNA Silencing: Methods and Protocols facilitates the translation of gene silencing concepts into practical applications, and includes a broad and useful set of RNA silencing protocols. Sections cover the biochemical aspects of silencing machinery, methods for RNA silencing in nonmammalian organisms, design, preparation, and use of RNAs to silence gene expression, several methos for the in vivo delivery of siRNAs and silencing vectors, and methods for the study and use of microRNAs.
Provides diverse, novel, and useful descriptions of miRNAs in several species, including plants, worms, flies, fish, chicks, mice, and humans. These include some useful adaptations and applications that could be relevant to the wider research community who are already familiar with the identification of miRNAs. This goal of this volume is to stimulate the reader to explore diverse ways to understanding the mechanism in which miRNAs facilitate the molecular aspects of the biomedical research.
Tested and proven solutions to the challenges of biologicaldrug product development Biological drug products play a central role in combating humandiseases; however, developing new successful biological drugspresents many challenges, including labor intensive productionprocesses, tighter regulatory controls, and increased marketcompetition. This book reviews the current state of the science,offering readers a single resource that sets forth the fundamentalsas well as tested and proven development strategies for biologicaldrugs. Moreover, the book prepares readers for the challenges thattypically arise during drug development, offering straightforwardsolutions to improve their ability to pass through all theregulatory hurdles and deliver new drug products to the market. Biological Drug Products begins with generalconsiderations for the development of any biological drug productand then explores the strategies and challenges involved in thedevelopment of specific types of biologics. Divided into fiveparts, the book examines: Part 1: General Aspects Part 2: Proteins and Peptides Part 3: Vaccines Part 4: Novel Biologics Part 5: Product Administration/Delivery Each chapter has been prepared by one or more leading experts inbiological drug development. Contributions are based on acomprehensive review and analysis of the current literature as wellas the authors' first-hand experience developing and testing newdrugs. References at the end of each chapter serve as a gateway tooriginal research papers and reviews in the field. By incorporating lessons learned and future directions forresearch, Biological Drug Products enables pharmaceuticalscientists and students to improve their success rate in developingnew biologics to treat a broad range of human diseases.
Long recognized as the standard general reference work providing a complete overview of contemporary gynaecological practice, this new edition of Shaw, Gynaecology provides all the information that trainees need to master in order to successfully take their professional certification exams as well as providingthe practicing gynaecologist with an accessible overview of the "state of play" of diagnostic and therapeutic procedures. Totally rewritten, it gives a succinct but comprehensive account of all currently available resources in the management of gynaecological disorders. Comprehensive overview of contemporary gynaecological practice with a clinically focused approach. It covers all of the areas that a gynaecologist covers on a day-to-day basis and helps in the formulation and implementation of the most effective treatment. Details the use of various imaging modalities and investigative techniques as they relate to specific diseases in order to provide a solid foundation for clinical practice. User-friendly features such as chapter outlines, summary tables, key point boxes incorporated throughout.Provides quick access to the most necessary information for practitioners needing a quick consult or trainees preparing for exams. Copiously illustrations clarify and enhance the text whenever appropriate. Highly selective and current list of references quickly directs the reader to further investigations. New full colour illustrations incorporated throughout to accurately depict the full range of both common and rare disorders. Details up-to-date investigative and minimally invasive therapeutic techniques to keep the user abreast of the latest diagnostic and management options. Enhanced emphasis on surgical outcomes to help the user select the most appropriate procedure for any given patient. Two brand new editors and many brand new contributors provide a fresh perspectives on gynaecological oncology, reproductive and urogynaecological conditions.