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This book spans diverse aspects of modified nucleic acids, from chemical synthesis and spectroscopy to in vivo applications, and highlights studies on chemical modifications of the backbone and nucleobases. Topics discussed include fluorescent pyrimidine and purine analogs, enzymatic approaches to the preparation of modified nucleic acids, emission and electron paramagnetic resonance (EPR) spectroscopy for studying nucleic acid structure and dynamics, non-covalent binding of low- and high-MW ligands to nucleic acids and the design of unnatural base pairs. This unique book addresses new developments and is designed for graduate level and professional research purposes.
A review of innovative tools for creative nucleic acid chemists that open the door to novel probes and therapeutic agents Nucleic acids continue to gain importance as novel diagnostic and therapeutic agents. With contributions from noted scientists and scholars, Enzymatic and Chemical Synthesis of Nucleic Acid Derivatives is a practical reference that includes a wide range of approaches for the synthesis of designer nucleic acids and their derivatives. The book covers enzymatic (including chemo-enzymatic) methods, with a focus on the synthesis and incorporation of modified nucleosides. The authors also offer a review of innovative approaches for the non-enzymatic chemical synthesis of nucleic acids and their analogs and derivatives, highlighting especially challenging species. The book offers a concise review of the methods that prepare novel and heavily modified polynucleotides in sufficient amount and purity for most clinical and research applications. This important book: -Presents a timely and topical guide to the synthesis of designer nucleic acids and their derivatives -Addresses the growing market for nucleotide-derived pharmaceuticals used as anti-infectives and chemotherapeutic agents, as well as fungicides and other agrochemicals. -Covers novel methods and the most recent trends in the field -Contains contributions from an international panel of noted scientistics Written for biochemists, medicinal chemists, natural products chemists, organic chemists, and biotechnologists, Enzymatic and Chemical Synthesis of Nucleic Acid Derivatives is a practice-oriented guide that reviews innovative methods for the enzymatic as well as non-enzymatic synthesis of nucleic acid species.
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Preface: In the past the chemical and enzymatic synthesis of oligonucleotides of defined sequence had to be left to a few experts. Now, however, with the triester approach, the phosphite method and the solid-support techniques gene fragment synthesis has turned into an easy procedure even for a non-chemist. Due to the elegant chemistry involved, all methods work without sophisticated equipment and are prone to mechanisation and eventual automation. It is hoped that combined chemical-encymatic gene synthesis may become a standard technique in a molecular biology laboratory, such as DNA sequencing or in-vitro recombination of nucleic acides. We omitted chemical RNA synthesis, since this field is developing so rapidly at the moment that one has to refer to the original publications. However, we included enzymatic synthesis of RNA fragments, procedures which already have obtained a high degree of standardisation. Most of the contributions are revised versions of the protocols supplied for the EMBO sponsored course on "Automated Chemical and Enzymic Gene Synthesis", held in Darmstadt, March 21 to April 3, 1982. The protocols were improved on the basis of the experience of 30 student scientists with chemical, biological or medical backgrounds. Previously omitted procedures, such as the wandering spot method for oligonucleotide analysis, were included. In editing the manuscript we encountered problems with the nomenclature of nucleic acid components. In unambiquous cases we favoured a simple description, hoping for example, that oligodeoxynucleotide is always understood to mean oligo-2'-deoxyribonucleotide. This book aims to provide those interested in DNA/RNA research with state-of-the-art methods in the synthesis, purification, and analysis of DNA and RNA fragments. The editors wish to thank the authors for their efforts in preparing manuscripts from the the revised laboratory protocols. We gratefully acknowledge the skill and the patience of Mrs. E. Ronnfeldt in typing the manuscripts. We express our thanks to Verlag Chemie for the friendly and very efficient cooperation.--H.G. Gassen A. Lang--Darmstadt, in July 1982.
This book features a special subsection of Nanomedicine, an application of nanotechnology to achieve breakthroughs in healthcare. It exploits the improved and often novel physical, chemical and biological properties of materials only existent at the nanometer scale. As a consequence of small scale, nanosystems in most cases are efficiently uptaken by cells and appear to act at the intracellular level. Nanotechnology has the potential to improve diagnosis, treatment and follow-up of diseases, and includes targeted drug delivery and regenerative medicine; it creates new tools and methods that impact significantly upon existing conservative practices. This volume is a collection of authoritative reviews. In the introductory section we define the field (intracellular delivery). Then, the fundamental routes of nanodelivery devices, cellular uptake, types of delivery devices, particularly in terms of localized cellular delivery, both for small drug molecules, macromolecular drugs and genes; at the academic and applied levels, are covered. The following section is dedicated to enhancing delivery via special targeting motifs followed by the introduction of different types of intracellular nanodelivery devices (e.g. a brief description of their chemistry) and ways of producing these different devices. Finally, we put special emphasis on particular disease states and on other biomedical applications, whilst diagnostic and sensing issues are also included. Intracellular delivery / therapy is a highly topical which will stir great interest. Intracellular delivery enables much more efficient drug delivery since the impact (on different organelles and sites) is intracellular as the drug is not supplied externally within the blood stream. There is great potential for targeted delivery with improved localized delivery and efficacy.
Thymidine analogue bearing a methyl ester at the C5 position were synthesized. Thermophilic family B DNA polymerases accepted these analogues as substrate to form the corresponding PCR product. Modified DNA containing this analogue were prepared by PCR with KOD Dash DNA polymerase and were further post-modified to form the corresponding derivatized DNA. These DNA could act as a template for PCR and could accept natural TTP or modified analogue as substrate. The thymidine analogue with an amino-linker was also reacted with fluorescein isothiocyanate, yielding a new analogue which was used as a substrate for PCR to get the corresponding fluorescent labeled DNA. Finally, a novel modified DNA aptamer for sialyllactose that is ubiquitous component of the cell surface responsible for the infections of several viruses was produced by using SELEX method with strong binding ability.