Polynucleotide synthesis labeling chemistry
Methods and compositions for making nucleoside phosphoramidites and nucleic acids, including mono-, di-, and polynucleotides, comprising a linker covalently attached to a levulinyl moiety are provided. A levulinyl-protected linking moiety affords an orthogonal approach to modifying a polynucleotide during or after solid phase synthesis with a molecule of interest, for example, a conjugate or a dye.
1. A composition, comprising:
wherein L comprises a linker; Z 1 is an oxygen or a sulfur; Z 2 is an oxygen or a sulfur; and S comprises a solid support useful for the synthesis of nucleic acids, wherein the linker is attached to S through a covalent linkage, and the linker comprises a functional group through which the linker can attach to a polynucleotide, and the linker further comprises at least one of an alkane, an alkene, a polyether, a polyalcohol, a polyester, a polyamine, a polyalkylamine, a polyphosphodiseter, a polyethylene glycol, a polypropylene, a propylene glycol, a mixture of ethylene and propylene glycols, or a combination thereof.
2. The composition according to claim 1 , wherein the linker moiety comprises an alkene, a polyether, a polyalcohol, a polyester, a polyamine, a polyalkylamine, a polyphosphodiester, or a combination thereof.
3. An oligonucleotide attached to a solid synthesis support, wherein the solid synthesis support comprises a composition of claim 1 .
4. An oligonucleotide attached to a solid synthesis support, wherein the solid synthesis support comprises a composition of claim 1 , and the oligonucleotide comprises one or more compositions comprising:
wherein B comprises a nitrogenous base that is modified or unmodified; L comprises a linker moiety; Z 1 is an oxygen or a sulfur; Z 2 is an oxygen or a sulfur; P 1 comprises a blocking group; and P 2 comprises a phosphorous moiety.
5. An oligonucleotide attached to a solid synthesis support, wherein the solid synthesis support comprises a composition of claim 1 , and the oligonucleotide comprises one or more compositions comprising:
wherein L comprises a non-nucleosidic linker moiety; Z 1 is an oxygen or a sulfur; Z 2 is an oxygen or a sulfur; and P comprises a phosphorous moiety.
6. An oligonucleotide attached to a solid synthesis support, wherein the solid synthesis support comprises a composition of claim 1 , and the oligonucleotide comprises one or more compositions comprising:
wherein B comprises a nitrogenous base that is modified or unmodified; L comprises a linker moiety; Z 1 is an oxygen or a sulfur; Z 2 is an oxygen or a sulfur; P 1 comprises a blocking group; and P 2 comprises a phosphorous moiety, and
a composition comprising:
wherein L comprises a non-nucleosidic linker moiety; Z 1 is an oxygen or a sulfur; Z 2 is an oxygen or a sulfur; and P comprises a phosphorous moiety.
7. A method of synthesizing an oligonucleotide comprising adding one more nucleotides to the composition of claim 1 .
8. The composition of claim 1 wherein the linker is attached to the solid support through a succinate, glutarate or oxalate linkage.
9. The composition of claim 1 , wherein the solid support comprises controlled pore glass, polystyrene or polymethacrylate.
10. The composition of claim 1 , wherein S and L are attached through a covalent amide bond.
11. The composition of claim 1 , wherein the covalent linkage is through a converted hydroxyl group.
12. The composition of claim 1 , wherein Z 1 is sulfur.
13. The composition of claim 1 , wherein Z 2 is sulfur.
14. The composition of claim 12 , wherein Z 2 is sulfur.