Oligonucleotide probe antisense

An alternative approach to clone the receptor was taken by Thomas and colleagues (1990, 1991). They screened a cDNA library from peritoneal rabbit cells with an antisense oligonucleotide probe deduced from the coding region of the second transmembrane domain, which is common to G-protein-coupled receptors (fMet-Leu-Phe is a G-protein-linked receptor). Screening of this library yielded a putative receptor clone F3R, characterised as follows ... 

In contrast to the biopharmaceuticals discussed thus far (recombinant proteins and gene therapy products), antisense oligonucleotides are manufactured by direct chemical synthesis. Organic synthetic pathways have been developed, optimized and commercialized for some time, as oligonucleotides are widely used reagents in molecular biology. They are required as primers, probes and for the purposes of site-directed mutagenesis. 

Scheme 2. Top, synthesis of a paramagnetic oligonucleotide, (i) DTPA dianhydride in DMSO (ii) GdCl3 bottom binding of the antisense probe to E. Coli 5 S rRNA [77]...

Lambert, R. C., Maulet, Y., Dupont, J. L., et al. Polyethylenimine-mediated DNA transfection of peripheral and central neurons in primary culture Probing Ca2+ channel structure and function with antisense oligonucleotides. Mol. Cell. Neurosci. 7 239-246, 1996. 

Hamon M, Doucet E, Lefevre K, et al. Antibodies and antisense oligonucleotide for probing the distribution and putative functions of central 5-HT6 receptors. Neuropsychopharmacology 1999 21 68S-76S. 

The DNA of a cell must be copied if the cell is to reproduce. Also, certain enzymes are required for a cell to metabolize and function (as contrasted with the dormant state), and these are usually made in the cell as needed. Therefore, an antisense probe that reacts with, for example, a critical RNA in the cell, may effectively render the cell dormant. One can envision this in vitro and in vivo. But oligonucleotides are subject to degradation by nucleases which, depending on their specificity, clip a nucleotide at certain base sequences,... 

DNA applications, in particular DNA sequencing, human identification, and genetic analysis dominate the field. Other DNA applications including oligonucleotides, antisense DNA, restriction fragments, plasmids, PCR products, hydridization (DNA probe), and RNA have also been demonstrated [7,8]. 

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