Nucleotide cleaving agents and method
AssigneeRegents of the University of Minnesota
InventorsLawrence Que, Jr., Richard S. Hanson, Leah M. T. Schnaith
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Abstract
The present invention provides a unique series of nucleotide cleaving agents and a method for cleaving a nucleotide sequence, whether single-stranded or double-stranded DNA or RNA, using and a cationic metal complex having at least one polydentate ligand to cleave the nucleotide sequence phosphate backbone to yield a hydroxyl end and a phosphate end.
Claims28
An in vitro method for cleaving a nucleotide sequence comprising contacting the nucleotide sequence to be cleaved with a cationic dinuclear metal complex with one or two polydentate ligands to cleave the nucleotide sequence in its phosphate backbone to form a hydroxyl end and a phosphate end, wherein the polydentate ligand is tethered to a nucleotide sequence recognition element.
An in vitro method for cleaving a nucleotide sequence comprising contacting the nucleotide sequence to be cleaved with a cationic dinuclear metal complex selected from the group of Fe.sub.2 (HPTB)(OH)(NO.sub.3).sub.4, Fe.sub.2 (HPTP)(OH)(NO.sub.3).sub.4, and [Fe.sub.2 (HPTP)(OBz)](BPh.sub.4).sub.2, wherein HPTB and HPTP are tethered to a nucleotide sequence recognition element.
An in vitro method for cleaving a supercoiled double-stranded DNA sequence comprising contacting the supercoiled double-stranded DNA sequence to be cleaved with a cationic dinuclear metal complex with one or two polydentate ligands to cleave the nucleotide sequence in its phosphate backbone to form a hydroxyl and a phosphate end on each original DNA strand, wherein the polydentate ligand is tethered to a nucleotide sequence recognition element.
An in vitro method for cleaving a nucleotide sequence comprising contacting the nucleotide sequence to be cleaved with a cationic dinuclear metal complex with one or two ligands, each containing two tridentate chelating groups linked by a polyatomic chain, to cleave the nucleotide sequence in its phosphate backbone to form a hydroxyl and a phosphate end;
- wherein:
- (a) the metal complex includes a metal selected from a group consisting of a Group IIIA, IB, VB, VIIB, VIIIB, or a lanthanide series metal;
- and
- (b) the ligand is represented by the formula (R.sup.1)(R.sup.2)N--R.sup.3 --N(R.sup.4)(R.sup.5) wherein:
- (I) each of R.sup.1, R.sup.2, R.sup.4, and R.sup.5 is independently a Lewis basic group;
- and
- (ii) R.sup.3 is a substituted or unsubstituted aromatic or aliphatic group with at least a 3 carbon chain;
- and
- wherein the polydentate ligand is tethered to a nucleotide sequence recognition element.
Dependencies are inferred from the claim text.