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(Chest. 2002;121:91S-97S.)
© 2002 American College of Chest Physicians

Mode of Action of RNA/DNA Oligonucleotides*

Progress in the Development of Gene Repair as a Therapy for {alpha}1-Antitrypsin Deficiency

Richard Metz, PhD; Mike DiCola, BS; Takao Kurihara, PhD; Austin Bailey, PhD; Bruce Frank, PhD; Bryan Roecklein, PhD and Michael Blaese, MD

* From ValiGen, Inc, Princeton, NJ.

Correspondence to: Richard Metz, 37 Winthrop Rd, Lawrenceville, NJ 08648; e-mail: janislmetz{at}aol.com

We describe a technology developed for the site-specific correction of a single base carried on an episome or chromosome in prokaryotic and eukaryotic cells. Critical to the development of this technology as a therapeutic device for treating genetic disorders, like {alpha}1-antitrypsin deficiency, is the establishment of a standardized assay to study its mode of action and structure-activity relationships (SARs). To this end, a positive-selection system in Escherichia coli has been developed to assess RNA/DNA oligonucleotide (RDO)-directed repair activity. We demonstrate that RDO-directed repair requires the concerted action of the two following repair proteins: the pairing protein RecA; and the mismatch recognition protein, MutS. SAR studies demonstrate that the RDO molecule is functionally asymmetric. The RNA-containing strand enables strand-pairing and stabilization of the molecule, and the DNA-containing strand confers the information transfer.




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V. T. Ciavatta, S. A. Padove, J. H. Boatright, and J. M. Nickerson
Mouse Retina Has Oligonucleotide-Induced Gene Repair Activity
Invest. Ophthalmol. Vis. Sci., July 1, 2005; 46(7): 2291 - 2299.
[Abstract] [Full Text] [PDF]


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