Antisense oligonucleotides methods

A selective method of preventing the expression of adhesion molecules or cytokines is the use of antisense oligonucleotides. These oligonucleotides are short sequences of nucleic acids complementary to mRNA sequences of specific proteins of interest. If delivered to the cytoplasmic compartment of cells these oligonucleotides are able to form a complex with their target mRNA. In this way the translation of mRNA into protein by ribosomes is inhibited. The subsequent mRNA degradation by RNAse H results in reduced expression of the protein (see also Chapter 5 for a description of antisense ohgonucleotides as therapeutic modalities). 

I. Doignon, F. Pillon, C. David, C. Crouzel, B. Tavitian, A general method to label antisense oligonucleotides with radioactive halogens for pharmacological and imaging studies, Bioconj. Chem. 11 (2000) 627-636. 

Zimmer, A. (1999). Antisense oligonucleotide delivery with polyhexylcyanoacrylate nanoparticles as carriers. Methods, 18(3), 286-295, 322. 

After a comprehensive introduction, the basic PK/PD properties of peptides, monoclonal antibodies, antisense oligonucleotides and gene delivery vectors are reviewed. In the second part, the book covers a number of challenges and opportunities in this field such as bioanalytical methods, bioequivalence and pulmonary delivery. The text finishes with a detailed presentation of some real-life examples and case studies which should be of particular interest to the reader and integrate many of the concepts presented earlier in the text. 

Since the first reports on magnetically enhanced nucleic acid delivery in the year 2000 (1,2), magnetofection has become a well-established method and has been predominantly used for in vitro applications. It has been shown to potentiate viral (3, 4) and non-viral nucleic acid delivery, including plasmids or small constructs such as antisense oligonucleotides, and synthetic siRNA and PCR products (5-10). The nucleic acids can be directly associated with magnetic nanoparticles in... 

Yu, R.Z., Geary, R.S., and Levin, A.A. (2004) Application of novel quantitative bioanalytical methods for pharmacokinetic and pharmacokinetic/pharmacodynamic assessments of antisense oligonucleotides. Current Opinion in Drug Discovery and Development, 7, 195 203. 

Chiron Corporation Oligonucleotide synthesis Screening method for antisense oligonucleotides development W09617955 1996... 

Zimmer A (1999). Antisense Oligonucleotide Delivery with Polyhexylcyanoacrylate Nanoparticles as Carriers. Methods. 18 286-295. 

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