In Oligonucleotide Synthesis: Methods and Applications, laboratory experts describe in step-by-step detail the powerful new techniques they have developed for oligonucleotide synthesis and for the use of modified oligonucleotides in ...
Author: Piet Herdewijn
Publisher: Springer Science & Business Media
Nucleic acids chemistry has progressed rapidly in the last 10 years with the introduction of many new techniques and the development of many novel therapeutic entities. In Oligonucleotide Synthesis: Methods and Applications, laboratory experts describe in step-by-step detail the powerful new techniques they have developed for oligonucleotide synthesis and for the use of modified oligonucleotides in biotechnology. Among the protocol highlights are the synthesis of phosphorothioates via new sulfur transfer agents, the synthesis of LNA, peptide conjugation methods to improve cellular delivery and cell-specific targeting, triple helix formation, and a novel two-step process that yields a high purity DNA that is less costly. Novel applications include the use of molecular beacons to monitor the PCR amplification process, nuclease footprinting to study the sequence-selective binding of small molecules to DNA, nucleic acid libraries, optimization of RNA synthesis for siRNA applications, and the use of small interference RNA (siRNA) as an inhibitor of gene expression. The protocols follow the successful Methods in Molecular BiologyTM series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and highly practical, Oligonucleotide Synthesis: Methods and Applications offers today's investigators not only insight into important new biotechnologies, but also a full range of the detailed protocols needed to work successfully in DNA synthesis and its applications today.
Oligonucleotide synthesis has become an essential technique required by many
biologically orientated laboratories . Yet up to now there has been no practical
manual available that adequately covers all the important techniques of synthetic
OLIGONUCLEOTIDES CHEMICAL SYNTHESIS CPG oligonucleotides with
modified termini and nicked dumbbell Structure show enhanced ... Novel base-
labile protecting groups for 5'-hydroxy function in solidphase oligonucleotide synthesis.
New photolabile protecting groups in nucleoside and nucleotide chemistry -
Synthesis , cleavage mechanisms and applications . Nucleosides & Nucleotides
17 , 1987 - 1996 ( 1998 ) . 75 . Gao , X . L . et al . Oligonucleotide synthesis using
The medicinal chemistry effort directed toward improving antisense and antigene oligonucleotides has synthesized a large ... p 5-18 In recent years there has been
an explosion of efi'ort devoted to the synthesis of oligonucleotide analogues.
Author: Derek J. Chadwick
Publisher: John Wiley & Sons
The use of oligonucleotides as therapeutic agents rests upon their ability to interfere, in a sequence-specific manner, with the fundamental machinery of protein synthesis either by binding to the mRNAs transcribed from a gene or by binding directly to a target gene. This approach can be used not only for inhibition of the synthesis of host proteins but also of those required by invading pathogens. Potential therapeutic applications are enormous, ranging over hypertension, cardiovascular disease, autoimmune disease, vital and other parasitic infections (especially HIV), and cancer. This book discusses the chemistry and pharmacokinetics of oligonucleotides and their analogues, and surveys the results of structure-activity studies and current clinical trials. It also critically reviews the problems with antisense therapy, such as the enzymatic destruction of oligonucleotides, the doses required for a therapeutic response, the difficulty in directing oligonucleotides to particular target tissues and cells, the need for parenteral administration, and doubts concerning the mechanism of action (especially problems associated with non-specific binding to proteins) and long-term effects.
The savings get much more dramatic as the number of syntheses is increased.21
B. Linkage between the First Nucleoside/Nucleotide and the Polymer Support
Almost all polymer-supported oligonucleotide synthesis is directed from the 3' ...
Hardbound. This volume is intended to cover the chemistry of one of the most widely studied and important natural products, DNA. Discussed in detail are physicochemical properties of the molecule itself as well as small-molecule natural products that are known to interact with it. Also included are methods to synthesize and manipulate DNA and modified analogues. Twenty chapters are devoted to this overall topic.The first five relate to the structure of DNA; the first focuses on thermodynamics and kinetics of double helix formation; the next two describe triple- and tetra- helical structures formed by DNA; and the last two focus on methods for probing DNA structure (specifically, NMR methods and chemical probing methods, respectively).Chapters 6-12 focus on the chemistry of natural DNA and modified analogues. The first of these addresses nonenzymatic methods for synthesizing DNA, and the next chapter, on methods for attachment of reporter groups t
2 and any of the oligonucleotide synthesis reagents . This means that both the
dimethyl ketal and 1 , 3 - dioxolane could potentially be used as a safety - catch
in our linker design since they are stable to the conditions / reagents of
2 APPLICATIONS OF OLIGONUCLEOTIDES The term oligonucleotide is often
used to mean specifically oligodeoxyribonucleotide , but this usage should be
avoided as methods for the synthesis of oligoribonucleotides are also now
While the use of 5 ' - phosphoramidites has been restricted primarily to the synthesis of 5 ' - 5 ' and 3 ' - 3 ' linked oligonucleotides , ” cently efforts that exploit
automated DNA synthesis in a 5 ' → 3 ' direction have gained momentum . DNA
Oligonucleotide synthesis v as low in dilute solutions of RNase A , rose steeply
as the enzyme concentration was increased , and reached a plateau at a
concentration of 40 ug per ml for Me - pUpU and pUpU . With U - cyclic - p the
Vols. 3-140 include the society's Proceedings, 1907-41
26 Letsinger and Mahadevan : Oligonucleotide Synthesis on a Polymer Support
27 Letsinger and Mahadevan : Stepwise Synthesis of Oligodeoxyribonucleotides
on an Insoluble Polymer Support 28 Letsinger , Caruthers , and Jerina ...
2 Chemical synthesis of oligonucleotides There are several methods of “ de novo
" oligonucleotide synthesis ; the dominant one is the phosphoramidite approach
developed by Caruthers and his coworkers in the early 1980s ( 4 , 5 , 6 ) .
28 OligoPrepTM : A New PVA Support for Oligonucleotide Synthesis at Large
Scale Zhiwei Wang , Isaiah Cedillo , Douglas L . Cole and Yogesh S . Sanghvi *
Development Chemistry , Isis Pharmaceuticals , Inc . , 2292 Faraday Avenue ...
The third approach entails the synthesis of oligonucleotide conjugates in solution
. ... in the 2 - position should be stable under conditions of oligonucleotide synthesis and be easily removed in the course of standard post - synthetic