FUNCTIONAL NUCLEIC ACIDS FOR BIOLOGICAL SEQUESTRATION

• US 20090266760A1
• Filed: 03/07/2008
• Published: 10/29/2009
• Est. Priority Date: 03/08/2007
• Status: Abandoned Application

• Current Assignee: BioTex Incorporated
• Original Assignee: BioTex Incorporated
• Inventors: Jackson, George William, McNichols, Roger Joseph
• Application Number:
  US12/044,737
• Publication Number:
  US 20090266760A1
• Time in Patent Office: Days
• US Class Current: 210/601
• CPC Class Codes: C12N 15/1048 SELEX
• CPC Class Codes: C12N 15/63 Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression

Abstract

The present invention generally relates to methods of improving the removal and/or treatment of substances in bulk volumes, particularly to methods of improving the removal and/or treatment of contaminants in bulk volumes by nucleic acid interaction and by including such nucleic acid interactions in organisms. The present invention further relates to methods for generating and/or improving the interaction of nucleic acids with substances for removal and/or treatment. Bulk volumes may generally refer to any volume of substance wherein the removal and/or treatment of substances therein may occur. Nucleic acids may be utilized to bind and/or catalytically interact with substances in the bulk volume. Further, the nucleic acids may be included in an organism for sequestering substances within cells.

Claims

1. An expression vector comprising:

a chimeric gene encoding selective nucleic acid ligands within a non-coding nucleic acid, operatively linked to a functional promoter, wherein said expression vector when transfected into a host transcribes said chimeric gene, and wherein said gene product is capable of binding to or altering at least one target molecule.

2. The expression vector of claim 1, wherein said vector further comprises at least one of a selection marker or a marker for selective induction.

3. The expression vector of claim 2, wherein said vector further comprises at least one selection marker and wherein said selection marker is an antibiotic resistance marker.

4. The expression vector of claim 1, wherein said promoter is a T7 RNA polymerase or a ribosomal RNA promoter.

5. The expression vector of claim 1, wherein said selective nucleic acid ligands are at least one of an aptamer, a ribozyme, an aptazyme or a riboswitch.

6. The expression vector of claim 1, wherein said non-coding nucleic acid is selected from the group consisting of rRNA, tRNA, RNAase P, small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), efference RNA (eRNA) and tmRNA.

7. The expression vector of claim 1, wherein said at least one target molecule is a waste fluid contaminant.

8. The expression vector of claim 7, wherein said waste fluid contaminant is at least one of an inorganic molecule, an organic molecule, a toxin, a peptide, or a viral particle.

9. The expression vector of claim 1, wherein said at least one target molecule is at least one of hormones, antibodies, proteins, enzymes, pharmaceuticals or metals.

10. An isolated cell comprising said expression vector of claim 1.

11. The cell of claim 10, wherein said cell is a prokaryotic cell or a eukaryotic cell.

12. An isolated cell comprising:

at least one nucleic acid ligand sequence incorporated into a genomic DNA encoding a non-coding nucleic acid, wherein said nucleic acid ligand sequence binds to or catalytically alters a target molecule.

13. The cell of claim 12, wherein said nucleic acid ligand sequence is incorporated into said non-coding nucleic acid by homologous recombination.

14. The cell of claim 12, wherein said non-coding nucleic acid is selected from the group consisting of rRNA, tRNA, RNAase P, small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), efference RNA (eRNA) and tmRNA.

15. The cell of claim 12, wherein said cell is a prokaryotic cell or a eukaryotic cell.

16. The cell of claim 12, wherein said nucleic acid sequence is an aptamer, a ribozyme, an aptazyme or a riboswitch.

17. The cell of claim 12, wherein said target molecule is a waste fluid contaminant.

18. The cell of claim 17, wherein said waste fluid contaminant is an inorganic molecule, an organic molecule, a toxin, a peptide, or a viral particle.

19. The cell of claim 12, wherein said target molecule is a hormone, an antibody, a protein, an enzyme, or a metal.

20. A method for sequestering within a cell a plurality of target molecules, present in a bulk volume comprising:

generating a library of nucleic acid ligand sequences capable of binding to said target molecules;

incorporating said nucleic acid ligand sequences into at least one non-coding nucleic acid within a cell;

culturing said cell to achieve a cell population;

contacting said cell population with said bulk volume; and

separating said cell population from said bulk volume.

21. The method of claim 20, further comprising recovering said target molecule from said cell population.

22. The method of claim 20, wherein said target molecules are at least one of an inorganic molecule, an organic molecule, a toxin, a peptide, a viral particle, a hormone, an antibody, a protein, an enzyme, a pharmaceutical or a metal.

23. The method of claim 20, wherein said separation is accomplished by a method selected from the group consisting of filtration, sedimentation, flocculation, adsorption, membrane filtration, biofilm formation and membrane bioreactor interaction.

24. The method of claim 20, wherein said non-coding nucleic acid is selected from the group consisting of rRNA, tRNA, RNAase P, small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), efference RNA (eRNA) and tmRNA.

25. The method of claim 20, wherein said incorporation of said nucleic acid ligand sequence is into said genomic DNA encoding said non-coding nucleic acid.

26. The method of claim 20, wherein said nucleic acid sequence is an aptamer, a ribozyme, an aptazyme or a riboswitch.

27. The method of claim 20, wherein said cell is a prokaryotic or a eukaryotic cell.

28. A method for bioremediation of at least one contaminant present in a bulk volume comprising:

generating a library of nucleic acid ligand sequences capable of binding to or altering said at least one contaminant;

incorporating said nucleic acid ligand sequences in at least one non-coding nucleic acid in a cell;

culturing said cell to achieve a cell population;

contacting said cell population with said bulk volume; and

,

separating said cell population from said bulk volume.

29. The method of claim 29, wherein said contaminants are at least one of an inorganic molecule, an organic molecule, a toxin, a protein, a peptide, and a viral particle.

30. The method of claim 29, wherein said bulk volume is at least one of bodily waste fluids, municipal waste water or effluent from an industrial process.