Method for biocatalytic protein-oligonucleotide conjugation
AssigneeThe Research Foundation for The State University of New York
InventorsBrian Callahan, Timothy Owen
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Abstract
A composition comprising a polypeptide ligated to an oligonucleotide through a sterol linker. A method of ligating a polypeptide to an oligonucleotide, comprising a polypeptide having a hedgehog steroyl transferase catalytic domain at the C-terminal of the polypeptide with an electrophilic residue, e.g., glycine, between polypeptide and the hedgehog steroyl transferase catalytic domain, and a steroylated oligonucleotide in solution, and permitting a reaction to cleave the hedgehog steroyl transferase catalytic domain from the polypeptide while ligating the steroylated oligonucleotide to the glycine at the C-terminal of the polypeptide. The oligonucleotide may be, for example, a therapeutic, diagnostic, or affinity ligand.
Claims19
A method for generating a polypeptide-nucleic acid conjugate, comprising:
- providing a polypeptide comprising a C-terminal steroyl transferase activity, an N-terminal peptide, and an intervening electrophilic residue;
- providing a nucleic acid substrate comprising a fused sterol or stanol ring system comprising:
- i) a nucleophilic substituent on the A ring;
- and
- ii) at least one nucleic acid, ligated to the fused sterol or stanol ring system by a linker;
- and
- reacting the nucleic acid substrate and the polypeptide in a solution, to form a covalently linked polypeptide-nucleic acid conjugate.
A method for ligating a polypeptide to a nucleic acid, comprising:
- providing a polypeptide comprising a C-terminal steroyl transferase activity, an N-terminal peptide, and an intervening electrophilic residue;
- providing a nucleic acid substrate for the steroyl transferase activity comprising a fused sterol or stanol ring system having a nucleophilic substituent of the A ring, the fused stanol or sterol system being ligated with a linker to at least one nucleic acid having an affinity for a complementary nucleic acid;
- and
- reacting the nucleic acid substrate and the polypeptide in a solution, to form a covalently linked polypeptide-nucleic acid conjugate.
Dependencies are inferred from the claim text.