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Peptide nucleic acids (PNA) were introduced in 1991 as a new class of pseudo-peptide DNA mimics conceived as ligands that recognize double-stranded DNA sequences specifically in the major groove. The present chapter outlines the different modes subsequently discovered for sequence-directed dsDNA targeting by PNA oligomers. This includes triple helix formation, triplex, duplex and double duplex invasion, and primarily focuses on efforts and progress made towards exploiting PNA for DNA-targeted control of gene expression in drug discovery approaches. In particular, recent in vivo animal studies are discussed.

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