Historical Perspective And Major Accomplishments Of Synthetic Oligonucleotides

The study of DNA originated with the work of Friedrich Miescher at the University of T bingen in Germany. Miescher was the first, in 1869, to isolate a new nonprotein substance, named nuclein, which was later to be identified as DNA.44 Although there was no indication back then that nuclein was a carrier of genetic information, Miescher was the first to realize that the contents of the nucleus had a unique chemical composition. His original studies were made using human pus cells, but he later...

Contents

Chemical Synthesis of Oligonucleotides From Dream to Artificial Hybridization Peptide Nucleic Acid (PNA) A DNA Mimic with a Pseudopeptide Synthetic DNA Used in Amplification Artificial Nucleic Acids and Genome Site-Directed Mimotopes An Artificial Genes for Chimeric Virus-Like DNA Sang Won Han, Jane Zveiter de Moraes, C lio Lopes Silva, Roger Chammas, and Mauricio Martins Rodrigues Toward Designer Diagnostic Recombinant

Ybu

Into highly reactive activated compounds (Figure 1.9) is one of the most remarkable aspects of phosphoramidite synthesis chemistry.223,234 This activation is performed by mixing a solution of nucleoside phosphoramidite with a solution of a very mild acid (activator). Initially, the activators were tertiary amine hydrochloride salts such as N,N-dimethylaniline hydrochloride, but since these reagents were hygroscopic and difficult to work with 1H-tetrazole soon became the preferred activator.102...

Info

Amino protecting groups that can be removed under mild or neutral conditions are also desirable for the synthesis of oligonucleotide analogues, which cannot withstand strongly basic cleavage conditions. The N-pent-4-enoyl NPT protecting group 48, which can be easily removed by treatment with either an iodine reagent r.t., 30 min , methanolic potassium carbonate r.t., 4 to 8 h , or ammonium hydroxide r.t., 15 min , is particularly noteworthy.160 This protecting group has been used to prepare...

H3c

The phenoxyacetyl 45 and the similar t-butylphenoxyacetyl 46 protecting group were introduced as more labile base protecting groups. These groups allowed the half-time for deprotection, using room temperature r.t. ammonium hydroxide, to be reduced to only 2 to 8 min. This fast depro-tection was particularly advantageous in the synthesis of oligoribonucleotides, which can undergo chain cleavage when treated with aqueous ammonium hydroxide for long periods of time.157 One minor problem with such...

Automated And Semiautomated Dna Synthesis

The primary advantage of solid-phase synthesis is the ease with which the solid-phase supports can be manipulated. Successful chemical strategies for oligonucleotide synthesis on solid-phase supports were a vast improvement over the tedious methods previously performed in solution. In the earliest work, solid-phase synthesis was manually performed using simple funnels or test tubes to hold the supports.104341 However, researchers sought easier and faster ways to perform the syntheses. One such...

Preface

The only approach to art is art itself. Klaus Eidam The True Life of Johann Sebastian Bach Research laboratories involved in studies on molecular genetic processes or on the development of diagnostic, prophylactic, and therapeutic tools for human diseases are all exceedingly familiar with the use of synthetic oligonucleotides or synthetic DNA. Today, the application of synthetic DNA is as common as the polymerase chain reaction PCR , DNA sequencing, or nucleic acid hybridization. What used to...

The Editors

Khudyakov, Ph.D., is Chief, Computational Molecular Biology Activity and Deputy Chief, Developmental Diagnostic Laboratory, Laboratory Branch, Division of Viral Hepatitis, National Center for Infectious Diseases NCID , Centers for Disease Control and Prevention CDC , Atlanta, Georgia. He received his M.Sc. in Genetics from the Novosibirsk State University, Novosibirsk, Russia in 1977 and his Ph.D. in Molecular Biology from the D.I. Ivanovsky Institute of Virology, Academy of Medical...