Gene expression with covalently modified polynucleotides
First Claim
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1. A complex for delivering nucleic acid to a cell, comprising:
- a plurality of a compound attached by a modifying chemical attachment to the nucleic acid in an amount sufficient to alter the tertiary structure of the nucleic acid when compared to the complex wherein the compound is not attached.
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Abstract
A process and compound wherein nucleic acids can be modified with a host of molecules and maintain their ability to be expressed. A modifying chemical attachment of polyions to polynucleotides can be used to facilitate the change of tertiary structure of the nucleic acid and in some cases condensation of nucleic acids into smaller, charged particles useful in delivering the nucleic acid to a cell.
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15 Claims
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1. A complex for delivering nucleic acid to a cell, comprising:
- a plurality of a compound attached by a modifying chemical attachment to the nucleic acid in an amount sufficient to alter the tertiary structure of the nucleic acid when compared to the complex wherein the compound is not attached.
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2. The complex of claim 18 wherein the change in tertiary structure results in a compaction or reduction in volume or a reduction in hydrodynamic radius of the nucleic acid.
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3. The complex of claim 18 wherein the change in tertiary structure results in a retention of greater than 40% expression of the expressible sequence when compared to unmodified nucleic acid.
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4. The complex of claim 18 wherein the compound attached to the nucleic acid is net positive, neutral, or net negative charge.
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5. The complex of claim 18 whereby the compound is attached to the N7 position of guanine.
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6. The complex of claim 18 wherein step of attaching comprises modifying the nucleic acid using an alkylating molecule.
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7. The complex of claim 23 wherein the alkylating molecule is selected from the group consisting of a mustard and a 3-membered ring system.
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8. The complex of claim 24 wherein the mustard is selected from the group consisting of a nitrogen mustard and a sulfur mustard.
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9. The complex of claim 25 wherein the 3-membered ring system is selected from the group consisting of aziridines, oxiranes, cyclopropyls, and episulfides.
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10. The complex of claim 26 wherein the nitrogen mustard consists of an R-chloride derivative.
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11. The complex of claim 24 wherein the 3-membered ring system consists of a CPI moiety.
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12. The complex of claim 18 wherein the step of attaching the compound comprises forming a Lewis acid:
- Lewis base complex, wherein the Lewis acid is not hydrogen.
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13. The complex of claim 29 wherein the Lewis acid is a transition metal.
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14. The complex of claim 30 wherein the Lewis acid is platinum.
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15. The complex of claim 29 wherein the Lewis base is N7 of guanine.
Specification