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Gene Targeting by Triple Helix-Forming Oligonucleotides
aLaboratory of Molecular Gerontology, National Institute on Aging, National Institutes of Health, Baltimore, Maryland 21224, USA bNovartis Pharmaceuticals, Horsham, West Sussex, RH12 4AB, United Kingdom cDepartment of Biochemistry and Molecular Biology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland 21205, USA
Address for correspondence: Michael M. Seidman, Laboratory of Molecular Gerontology, National Institute on Aging, National Institutes of Health, 5600 Nathan Shock Dr., Baltimore MD 21224. Voice: 410-558-8565; fax 410-558-8157. seidmanm{at}grc.nia.nih.gov Ann. N.Y. Acad. Sci. 1002: 141-153 (2003).
Effective gene targeting reagents would have widespread utility for genomic manipulation including transgenic cell and animal construction and for gene therapy. They would also be useful in basic research as probes of chromatin structure, and as tools for studying the repair and mutagenesis of targeted DNA damage. We are developing triple helix-forming oligonucleotides (TFOs) for gene targeting in living mammalian cells. Challenges to TFO bioactivity include the impediments to the biochemistry of triplex formation presented by the physiological environment and the charge repulsion between the duplex and the third strand. In addition, there are biological constraints to target access imposed by mammalian chromatin structure. Here we describe the oligonucleotide modification format that appears to support biological activity of TFOs. In addition we show that manipulation of the cell biology, specifically the cell cycle, has a dramatic influence on TFO bioactivity.
Key Words: DNA triple helix • gene targeting • psoralen crosslinking • cell cycle This article has been cited by other articles:
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