DNA and RNA Cleavage Mediated by Phenanthroline-Cuprous Oligonucleotides: From Properties to Applications

François, Jean‐Christophe; Faria, Marcella; Perrin, David M.; Giovannangéli, Carine

doi:10.1007/978-3-642-18510-6_13

Cited by 8 publications

(4 citation statements)

References 67 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Consequently, an LNA-containing TFO (TFO/ LNA) strongly bound to its chromosomal DNA target in a sequence-specific manner (22) and inhibited transcription at its target site in cultured cells, whereas a nonmodified TFO could not. Previous studies by our lab and others have shown that conjugating a molecule with DNA cleavage activity to a TFO results in sequence-specific cleavage in vitro (23), but to our knowledge, no targeted cleavage activity has been reported in cells. By conjugating orthophenanthroline (OP) to TFO/LNAs, we show here that the resulting OP-TFO/LNA conjugates are highly specific endonucleases in the chromosomal context and that they can be used for efficient targeted gene modification in human cells.…”

mentioning

confidence: 64%

RETRACTED: Triplex-forming oligonucleotide–orthophenanthroline conjugates for efficient targeted genome modification

Cannata

Brunet

Perrouault

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

The inefficiency of gene modification by homologous recombination can be overcome by the introduction of a double-strand break (DSB) in the target. Engineering the endonucleases needed, however, remains a challenging task that limits widespread application of nuclease-driven gene modification. We report here that conjugates of orthophenanthroline (OP), a DNA cleaving molecule, and triplex-forming oligonucleotides (TFOs), known to bind specific DNA sequences, are synthetic nucleases efficient at stimulating targeted genome modification. We show that in cultured cells, OP-TFO conjugates induce targeted DSBs. An OP-TFO with a unique target was highly efficient, and mutations at the target site were found in ≈10% of treated cells, including small deletions most likely introduced during DSB repair by nonhomologous end joining. Importantly, we found that when homologous donor DNA was cotransfected, targeted gene modification took place in >1.5% of treated cells. Because triplex-forming sequences are frequent in human and mouse genes, OP-TFO conjugates therefore constitute an important class of site-specific nucleases for targeted gene modification. Harnessing DNA-damaging molecules to predetermined genomic sites, as achieved here, should also provide inroads into mechanisms of DNA repair and cancer.

show abstract

mentioning

confidence: 64%

RETRACTED: Triplex-forming oligonucleotide–orthophenanthroline conjugates for efficient targeted genome modification

Cannata

Brunet

Perrouault

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…They have already been used to interfere intracellularly with processes occurring on DNA, such as transcription, repair, or recombination. Manipulation of chromatin structure (36, 37) and positioning of DNA-damaging agents using these agents conjugated with TFO (24,38) are other promising applications of DNA codereading molecules; they have been successfully explored with TFOs, but mainly in vitro. However, today, the major limitation of triplex-based applications is the limited intracellular stability of the complex.…”

Section: Discussionmentioning

confidence: 99%

Exploring Cellular Activity of Locked Nucleic Acid-modified Triplex-forming Oligonucleotides and Defining Its Molecular Basis

Brunet

Alberti

Perrouault

et al. 2005

Journal of Biological Chemistry

View full text Add to dashboard Cite

Triplex-forming oligonucleotides (TFOs), as DNAbinding molecules that recognize specific sequences, offer unique potential for the understanding of processes occurring on DNA and associated functions. They are also powerful DNA recognition elements for the positioning of ubiquitous molecules acting on DNA, such as anticancer drugs. A prerequisite for further development of DNA code-reading molecules including TFOs is their ability to form a complex in a cellular context: their binding affinities must be comparable to those of DNA-associated proteins. To reach this goal, chemically modified TFOs must be developed. In this work, we present triplex-forming properties (kinetics and thermodynamics) and cellular activity of G-containing locked nucleic acid-modified TFOs (TFO/LNAs). In conditions simulating physiological ones, these TFO/LNAs strongly enhanced triplex stability compared with the non-modified TFO or with the pyrimidine TFO/LNA directed against the same oligopyrimidine⅐oligopurine sequence, mainly by decreasing the dissociation rate constant and conferring an entropic gain. We provide evidence of their biological activity by a triplex-based mechanism, in vitro and in a cellular context, under conditions in which the parent phosphodiester oligonucleotide did not exhibit any inhibitory effect.The design of synthetic non-protein molecules able to control DNA-associated biological functions through their interaction with a specific DNA sequence represents a very attractive approach. Among DNA code-reading molecules, triplex-forming oligonucleotides (TFOs) 1 are able to bind to the major groove of oligopyrimidine⅐oligopurine regions in doublestranded DNA. A series of results have validated triplex-based approaches at the molecular and cellular levels: triplexes have been shown to interfere with transcription (initiation and elongation), replication, repair, and recombination (1, 2). However, the intracellular efficiency of TFOs still has to be improved.One possible approach consists of increasing the stability of the non-covalent triple helices under physiological conditions to reach binding affinities comparable to those of DNA-associated regulatory proteins. To this end, a variety of chemically modified nucleic acids have been developed. Among them are locked nucleic acids (LNAs) that contain LNA nucleotide monomers, i.e. ribonucleotides with a 2Ј-O,4Ј-C-methylene linkage that effects conformational fixation of the furanose ring in a C3Ј-endo conformation (3, 4). LNA-containing oligonucleotides have been recently shown to enhance triplex stability and to alleviate in part the sequence constraints imposed by the triple helical recognition motifs (5, 6). Only a few hybridization properties of LNA-modified TFOs (TFO/LNAs) have been reported so far, and they all concern (T,C)-containing TFO/LNAs. It has been shown that fully modified TFO/LNAs failed to bind to double-stranded DNA (7, 8), likely due to conformational restraint of TFO/LNA. However, alternating DNA and LNA nucleotides in TFO sequences is appropri...

show abstract

“…The 1,10-phenanthroline (phen) and its derivatives exhibit antiviral, antifungal and antimycoplasmal activities [ 10 ]. It has been reported that stable bis-phen complexes inhibited DNA or RNA polymerase activities and induced strand scission of DNA in the presence of reducing agents (thiol or H 2 O 2 ) [ 4 , 11 , 12 , 13 , 14 , 15 ]. DNA damage in the presence of Cu-phenanthrolines is attributed to the highly reactive hydroxyl radicals, OH· generated through the site-specific Fenton reaction [ 16 ].…”

Section: Introductionmentioning

confidence: 99%

Synthesis, Crystal Structure, Spectroscopic Properties and Potential Biological Activities of Salicylate‒Neocuproine Ternary Copper(II) Complexes

et al. 2015

View full text Add to dashboard Cite

Mixed ligand copper(II) complexes containing derivatives of salicylic acid and heterocyclic ligands with nitrogen donor atoms have been the subject of various studies and reviews. In this paper, synthesis and characterization of the ternary copper(II) complexes of neocuproine (2,9-dimethyl-1,10-phenanthroline, Neo) and salicylate ligands (Sal) are reported. In addition, the crystal structures of ([Cu(H2O)(5-Cl-Sal)(Neo)] (1), [Cu(μ-Sal)(Neo)]2 (2), Cu2(μ-5-Cl-Sal)(5-Cl-HSal)2(Neo)2]·EtOH (3)) were determined. In order to compare structural and biological properties of the prepared complexes, spectroscopic and biological studies were performed. Results of X-ray diffraction show that prepared complexes form three types of crystal structures in a given system: monomeric, dimeric and dinuclear complex. The preliminary study on the DNA cleavage activity has shown that the complexes under study behave as the chemical nucleases in the presence of added hydrogen peroxide with slight differences in the activity (1 > 2 > 3). The complexes 1 and 2 exhibited nuclease activity itself indicating the interaction of complexes with the DNA. It has been proposed that the enhanced destructive effect of the complexes 1 and 2 on the DNA is a result of two possible mechanisms of action: (i) the conversion of closed circular DNA (form I) to the nicked DNA (form II) caused by the copper complex itself and (ii) damage of DNA by Reactive Oxygen Species (ROS)—products of the interaction of copper with hydrogen peroxide by means of Fenton reaction (hydroxyl radicals). Thus the biological activity of the prepared Cu(II) complexes containing derivatives of salicylic acid and phenanthroline molecules is substantiated by two independent mechanisms. While derivatives of salicylic acids in the coordination sphere of copper complexes are responsible for radical-scavenging activity (predominantly towards superoxide radical anion), the presence of chelating ligand 2,9-dimethyl-1,10-phenanthroline significantly enhances capability of Cu(II) complexes binding to DNA via intercalation.

show abstract

DNA and RNA Cleavage Mediated by Phenanthroline-Cuprous Oligonucleotides: From Properties to Applications

Cited by 8 publications

References 67 publications

RETRACTED: Triplex-forming oligonucleotide–orthophenanthroline conjugates for efficient targeted genome modification

RETRACTED: Triplex-forming oligonucleotide–orthophenanthroline conjugates for efficient targeted genome modification

Exploring Cellular Activity of Locked Nucleic Acid-modified Triplex-forming Oligonucleotides and Defining Its Molecular Basis

Synthesis, Crystal Structure, Spectroscopic Properties and Potential Biological Activities of Salicylate‒Neocuproine Ternary Copper(II) Complexes

Contact Info

Product

Resources

About