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"A series of studies on 2'-fluorinated and 4'-chalcogen-modified nucleic acids is described, mainly directed toward the development of better gene silencing therapeutics. NMR/MD and osmotic stressing were used to compare the structure and hydration of 10-bp 2'F-ANA:RNA, ANA:RNA and DNA:RNA duplexes. The 2'F-ANA and ANA strands both featured sugars that pucker in the east (O4'-endo) conformation, as previously observed for hairpin structures containing hybrid stems. Osmotic stressing suggested that the 2'F-ANA:RNA duplex liberated fewer molecules of water upon melting than did ANA:RNA, which may give the former an entropic advantage that contributes to its far greater thermal stability. The stability of 2'F-ANA to acid-mediated hydrolysis was compared to that of DNA and RNA. Several phosphodiester (PO) or phosphorothioate (PS) 2'F-ANA sequences were incubated at pH ~1.2, and virtually no cleavage was observed after 2 days. In contrast, rapid degradation was observed for DNA (t1/2 = minutes) and RNA (t1/2 = hours (PO) or days (PS)). The nuclease-catalyzed hydrolysis of 2'F-ANA was also explored in detail. One diastereomer of the PS-2'F-ANA linkage was much more vulnerable to enzymatic cleavage than the other, which is parallel to the properties observed for PS-DNA. We also show that the nuclease stability induced by 2'F-ANA depends on the oligonucleotide sequence. An improved synthesis of 2'-deoxy-2'-fluoro-5-methyl-4'-thioarabinouridine (4'S-FMAU) is described. Participation of the 3'-O-benzoyl protecting group in the thiosugar precursor influenced the stereochemistry of the N-glycosylation reaction in nonpolar solvents, permitting a higher beta/alpha ratio than previously observed for similar Lewis acid-catalyzed glycosylations. The nucleoside adopted a predominantly northern conformation, in contrast to 2'-deoxy-2'-fluoro-5-methylarabinouridine (FMAU), which adopts a predominantly southeast conform"--
This hands-on guide to RNA interference brings the power of targeted gene silencing to any laboratory with the basic equipment for handling nucleic acids. In easy-to-follow, step-by-step protocols you will learn * how RNAi works in worms, flies and mammals, * how to design the most efficient RNAi constructs, * how to achieve transient, stable and conditional RNAi in cell cultures, * how to determine the efficiency of an RNAi experiment, * and how to use RNAi for gene therapy. All the protocols have been thoroughly tested in the author's own laboratory, and she provides examples of successful experiments and troubleshooting hints to help in establishing your own successful RNAi experiments. Also includes a list of suppliers for RNAi reagents and equipment as well as a glossary of terms.
This book focuses on various aspects of nanotechnology based gene silencing. The initial chapters detail the techniques available for in vitro and in vivo characterization of nanoparticles. In the later chapters, exhaustive details about various polymeric systems employed for gene silencing has been accounted.
In the past few decades there has been incredible growth in "bionano"-related research, which has been accompanied by numerous publications in this field. Although various compilations address topics related to deoxyribonucleic acid (DNA) and protein, there are few books that focus on determining the structure of ribonucleic acid (RNA) and using RNA as building blocks to construct nanoarchitectures for biomedical and healthcare applications. RNA Nanotechnology is a comprehensive volume that details both the traditional approaches and the latest developments in the field of RNA-related technology. This book targets a wide audience: a broad introduction provides a solid academic background for students, researchers, and scientists who are unfamiliar with the subject, while the in-depth descriptions and discussions are useful for advanced professionals. The book opens with reviews on the basic aspects of RNA biology, computational approaches for predicting RNA structures, and traditional and emerging experimental approaches for probing RNA structures. This section is followed by explorations of the latest research and discoveries in RNA nanotechnology, including the design and construction of RNA-based nanostructures. The final segment of the book includes descriptions and discussions of the potential biological and therapeutic applications of small RNA molecules, such as small/short interfering RNAs (siRNAs), microRNAs (miRNAs), RNA aptamers, and ribozymes.
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.
​The series Topics in Current Chemistry Collections presents critical reviews from the journal Topics in Current Chemistry organized in topical volumes. The scope of coverage is all areas of chemical science including the interfaces with related disciplines such as biology, medicine and materials science. The goal of each thematic volume is to give the non-specialist reader, whether in academia or industry, a comprehensive insight into an area where new research is emerging which is of interest to a larger scientific audience. Each review within the volume critically surveys one aspect of that topic and places it within the context of the volume as a whole. The most significant developments of the last 5 to 10 years are presented using selected examples to illustrate the principles discussed. The coverage is not intended to be an exhaustive summary of the field or include large quantities of data, but should rather be conceptual, concentrating on the methodological thinking that will allow the non-specialist reader to understand the information presented. Contributions also offer an outlook on potential future developments in the field. The chapter "Polymeric Nanoparticle-Mediated Gene Delivery for Lung Cancer Treatment" is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.
Gene Delivery into Mammalian Cells: An Overview on Existing Approaches Employed In Vitro and In Vivo, by Peter Hahn and Elizabeth Scanlan * Strategies for the Preparation of Synthetic Transfection Vectors, by Asier Unciti-Broceta, Matthew N. Bacon, and Mark Bradley * Cationic Lipids: Molecular Structure/Transfection Activity Relationships and Interactions with Biomembranes, by Rumiana Koynova and Boris Tenchov * Hyperbranched Polyamines for Transfection, by Wiebke Fischer, Marcelo Calderon, and Rainer Haag * Carbohydrate Polymers for Nonviral Nucleic Acid Delivery, by Antons Sizovs, Patrick M. McLendon, Sathya Srinivasachari, and Theresa M. Reineke * Cationic Liposome–Nucleic Acid Complexes for Gene Delivery and Silencing: Pathways and Mechanisms for Plasmid DNA and siRNA, by Kai K. Ewert, Alexandra Zidovska, Ayesha Ahmad, Nathan F. Bouxsein, Heather M. Evans, Christopher S. McAllister, Charles E. Samuel, and Cyrus R. Safinya * Chemically Programmed Polymers for Targeted DNA and siRNA Transfection, by Eveline Edith Salcher and Ernst Wagner * Photochemical Internalization: A New Tool for Gene and Oligonucleotide Delivery, by Kristian Berg, Maria Berstad, Lina Prasmickaite, Anette Weyergang, Pål K. Selbo, Ida Hedfors, and Anders Høgset * Visualizing Uptake and Intracellular Trafficking of Gene Carriers by Single-Particle Tracking, by N. Ruthardt and C. Bräuchle
The sequencing of the human genome and subsequent elucidation of the molecular pathways that are important in the pathology of disease have provided unprecedented opportunities for the development of new therapeutics. Nucleic acid-based drugs have emerged in recent years to yield extremely promising candidates for drug therapy to a wide range of diseases. Advances in Nucleic Acid Therapeutics is a comprehensive review of the latest advances in the field, covering the background of the development of nucleic acids for therapeutic purposes to the array of drug development approaches currently being pursued using antisense, RNAi, aptamer, immune modulatory and other synthetic oligonucleotides. Nucleic acid therapeutics is a field that has been continually innovating to meet the challenges of drug discovery and development; bringing contributions together from leaders at the forefront of progress, this book depicts the many approaches currently being pursued in both academia and industry. A go-to volume for medicinal chemists, Advances in Nucleic Acid Therapeutics provides a broad overview of techniques of contemporary interest in drug discovery.
Maximizing the potential of RNA interference in functional genomics - as well as in the development of therapeutics - continues to be at the forefront of biomedical research. Unlike journal articles, Gene Silencing by RNA Interference: Technology and Application combines essential background to the RNAi field with practical techniques designed by r