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Nucleic Acids Research, Vol 24, Issue 24 4924-4932, Copyright © 1996 by Oxford University Press

ARTICLES

In vivo persistence of DNA triple helices containing psoralen- conjugated oligodeoxyribonucleotides

M Musso, JC Wang and MW Van Dyke
Department of Tumor Biology, The University of Texas M. D. Anderson Cancer Center, Houston 77030, USA.

Triple helices represent an attractive method for modulating specific gene expression. In particular, cross-linking between a triplex-forming oligonucleotide (TFO) and its duplex DNA target, typically through the formation of psoralen photoadducts, allows efficient blocking of elongation by RNA polymerases in vitro. However, in vivo, this approach is limited by DNA repair of the photoadduct. Here we describe the use of an oligodeoxyribonucleotide 19mer psoralen-modified TFO to form covalent linkages between an oligonucleotide and both strands of the targeted duplex DNA, thereby efficiently blocking expression of a luciferase reporter gene. Most importantly, we demonstrate that both the psoralen cross-link and the purine-motif triplex remained intact for at least 72 h post-transfection, indicating that such species can persist for an extended period of time in vivo. These findings support the feasibility of an antigene approach for the therapeutic regulation of specific gene expression.
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