title: Polyamide fluorescent probes for visualization of repeated DNA sequences in living cells.
abstract: DNA imaging in living cells usually requires transgenic approaches that modify the genome. Synthetic pyrrole-imidazole polyamides that bind specifically to the minor groove of double-stranded DNA (dsDNA) represent an attractive approach for in-cell imaging that does not necessitate changes to the genome. Nine hairpin polyamides that target mouse major satellite DNA were synthesized. Their interactions with synthetic target dsDNA fragments were studied by thermal denaturation, gel-shift electrophoresis, circular dichroism, and fluorescence spectroscopy. The polyamides had different affinities for the target DNA, and fluorescent labeling of the polyamides affected their affinity for their targets. We validated the specificity of the probes in fixed cells and provide evidence that two of the probes detect target sequences in mouse living cell lines. This study demonstrates for the first time that synthetic compounds can be used for the visualization of the nuclear substructures formed by repeated DNA sequences in living cells.
authors: Structure and Instability of Genomes, Sorbonne Universités, Muséum national d'Histoire naturelle, INSERM U 1154, CNRS UMR 7196, 57 rue Cuvier, C.P. 26, 75231 Paris Cedex 05 (France).
authors: Karine
authors: K
authors: Nozeret
authors: François
authors: F
authors: Loll
authors: Christophe
authors: C
authors: Escudé
authors: Alexandre S
authors: AS
authors: Boutorine
mesh_terms: D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D051379:Mice; D008856:Microscopy, Fluorescence; D041681:NIH 3T3 Cells; D009757:Nylons / Q000737:chemistry*; D061848:Optical Imaging; D012091:Repetitive Sequences, Nucleic Acid
chemical_list: D005456:Fluorescent Dyes; D009757:Nylons; D004247:DNA
keywords: DNA recognition; bioorganic chemistry; fluorescent probes; live cells; minor-groove binders