Peptide Nucleic Acids: Methods and Protocols

ISBN-13: 9781617372858 / Angielski / Miękka / 2010 / 274 str.

Peter E. Nielsen

Opis

Peptide nucleic acids (PNAs) have now existed for slightly more than ten years, with the interest in and applications of this pseudopeptide DNA mimic steadily increasing during the entire period. PNAs have rapidly attracted the attention of scientists from a diversity of fields ranging from (bio)organic and biophysical chemistry to prebiotic evolution, and from molecular biology to genetic diagnostics and drug development. Many of the applications take advantage of the unique properties of PNA an uncharged pseudopeptide that distinguish this DNA mimic from more traditional DNA analogs. Rather than trying to create a comprehensive collection of all published methods and protocols involving PNA many of which have not yet been validated I have decided to concentrate on select protocols that are either very well established by several groups around the world, such as PCR-clamping and in situ hybridization, or on new methods that may have broader future impact. Basic methods for PNA oligomer synthesis and analyses have also been included. I am very grateful to those friends and colleagues who have enthusiastically contributed their work, discussions, and writing, and thereby made this book possible. Peter E. Nielsen v Contents Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix IINTRODUCTION 1 PNA Technology Peter E. Nielsen. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 II CHEMISTRY 2 Solid Phase Synthesis of PNA Oligomers Frederik Beck. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 3 Synthesis of PNA-Peptide Conjugates Satish Kumar Awasthi and Peter E. Nielsen. . . . . . . . . . . . . . . . . . 43 4 Parallel Synthesis of PNA-Peptide Conjugate Libraries Satish Kumar Awasthi and Peter E. Nielsen. . . . . . . . . . . . . . . . . ."

Recenzje

"The book, Peptide Nucleic Acid, provides an excellent introduction to pNA's...provides useful information for the readers in the development of delivery systems for bioactive macromolecules." -Pharmaceutical Research

Spis treści

Part I. Introduction PNA Technology Peter E. Nielsen Part II. Chemistry Solid Phase Synthesis of PNA Oligomers Frederik Beck Synthesis of PNA-Peptide Conjugates Satish Kumar Awasthi and Peter E. Nielsen Parallel Synthesis of PNA-Peptide Conjugate Libraries Satish Kumar Awasthi and Peter E. Nielsen Thermodynamics of PNA Interactions with DNA and RNA Tommi Ratilainen and Bengt Nordén Part III. PNA Technology Measurement of PNA Binding to Double-Stranded DNA Thomas Bentin, Georg I. Hansen, and Peter E. Nielsen PNA-Mediated Immobilization of Supercoiled DNA Thomas Bentin and Peter E. Nielsen PNA Openers and Their Applications Vadim V. Demidov and Maxim D. Frank-Kamenetskii Colorimetric Detection of PNA-DNA Hybridization Using Cyanine Dyes Miaomiao Wang and Bruce A. Armitage Part IV. Diagnostics PNA-Mediated PCR Clamping: Applications and Methods Deborah G. Murdock and Douglas C. Wallace PNA Clamping Techniques for the Determination of Oncogene Mutations Marcus Schuermann and Moira Behn. PNA Fluorescent In Situ Hybridization for Rapid Microbiology and Cytogenetic Analysis Brett Williams, Henrik Stender, and James M. Coull Detection of Point Mutations Using PNA-Containing Electrophoresis Matrices Gabor L. Igloi Part V. Antisense and Antigene Lipid-Mediated Introduction of Peptide Nucleic Acids Into Cells Dwaine A. Braasch and David R. Corey Synthesis of Cell-Penetrating Peptide-PNA Constructs Margus Pooga, Ursel Soomets, Tamas Bartfai, and Ülo Langel Antisense Inhibition of Bacterial Gene Expression and Cell Growth Liam Good In Vitro Transcription from Peptide Nucleic Acid/DNA Strand Displacement Loops Niels Erik Møllegaard and Peter E. Nielsen Part VI.Animal Studies In Vitro and in Vivo Studies on Pharmacokinetics and Metabolism of PNA Constructs in Rodents Edward Kristensen Index

Blurb

Peptide nucleic acid (PNA) technology continues to grow in importance throughout molecular biology, genetic diagnostics, and molecular medicine. In Peptide Nucleic Acids: Methods and Protocols, Peter Eigil Nielsen has assembled a critically evaluated collection of PNA protocols that are either already well established around the world, such as PCR-clamping and in situ hybridization, or that display promise of significant future impact. Basic methods for PNA oligomer synthesis and analysis have also been included. Written by experts with hands-on experience in the methods they describe, these readily reproducible protocols contain detailed step-by-step instructions, tips on avoiding pitfalls and on extending the method to other situations, and introductory material explaining the theory behind the process.
Up-to-date and highly practical, Peptide Nucleic Acids: Methods and Protocols provides for both novice and experienced researchers a thorough and easy introduction to the optimal use of today's PNA technology across a wide variety of fields, including molecular biology, medicinal chemistry, genetic diagnostics, and drug development.