Double duplex invasion by peptide nucleic acid: a general principle for sequence-specific targeting of double-stranded DNA.
Pseudocomplementary PNAs containing diaminopurine.thiouracil base pairs have been prepared and are shown to bind with high specificity and efficiency to complementary targets in double-stranded DNA by a mechanism termed "double duplex invasion" in which the duplex is unwound and both DNA strands are targeted simultaneously, each by one of the two pseudocomplementary peptide nucleic acids (PNAs). On the basis of our results we predict that (for decameric targets) more than 80% of all sequences can be targeted by straightforward Watson-Crick base pairing by using this approach in its present form. Targeting of pseudocomplementary PNAs to the promoter of the T7 phage RNA polymerase effectively inhibits transcription initiation. These results have important implications in the development of gene therapeutic agents as well as for genetic diagnostic and molecular biology applications.
Lohse
J
J
Center for Biomolecular Recognition, Department of Chemistry, University of Copenhagen, Universitetsparken 5, DK-2100 Copenhagen O, Denmark.
Dahl
O
O
Nielsen
P E
PE
D000225:Adenine / Q000737:chemistry; D001483:Base Sequence; D004247:DNA / Q000235:genetics* / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009696:Nucleic Acids / Q000378:metabolism*; D009841:Oligonucleotides / Q000235:genetics / Q000378:metabolism; D020135:Peptide Nucleic Acids; D010455:Peptides / Q000378:metabolism*; D011687:Purines / Q000737:chemistry; D013889:Thiouracil / Q000737:chemistry; D014158:Transcription, Genetic
D009696:Nucleic Acids; D009841:Oligonucleotides; D020135:Peptide Nucleic Acids; D010455:Peptides; D011687:Purines; D013889:Thiouracil; D004247:DNA; D000225:Adenine