Preparation and nuclease activity of hybrid "metallotris(methylpyridinium)porphyrin oligonucleotide" molecules having a 3'-loop for protection against 3'-exonucleases.
A 5'-GCGAAAGC minihairpin structure was added to the 3'-end of an oligonucleotide substituted at the 5'-end by a manganese cationic porphyrin in order to enhance the 3'-exonuclease resistance of these cleaver-antisense molecules. The influence of this minihairpin on the 3'-exonuclease resistance, the binding affinity to a target ssDNA, and the cleaving efficiency of Mn-cationic porphyrin oligonucleotide conjugates was compared to that of the parent molecule without the 3'-hairpin. The results showed that the 3'-hairpin slightly decreased the binding affinity and consequently the cleaving efficiency of the conjugated molecule toward a target sequence, but the much higher nuclease resistance makes 3'-minihairpin-protected metalloporphyrin oligonucleotides good candidates as reactive antisense oligonucleotides for studies on cells.
Mestre
B
B
Laboratoire de Chimie de Coordination du CNRS, Toulouse, France.
Pratviel
G
G
Meunier
B
B
D001483:Base Sequence; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000378:metabolism; D043211:Exodeoxyribonuclease V; D005090:Exodeoxyribonucleases / Q000378:metabolism*; D007202:Indicators and Reagents; D008345:Manganese; D008665:Metalloporphyrins; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis / Q000737:chemistry*; D013379:Substrate Specificity; D013816:Thermodynamics
D004277:DNA, Single-Stranded; D007202:Indicators and Reagents; D008665:Metalloporphyrins; D009838:Oligodeoxyribonucleotides; D016376:Oligonucleotides, Antisense; D008345:Manganese; D005090:Exodeoxyribonucleases; D043211:Exodeoxyribonuclease V