Plant transactivation interaction motifs and uses thereof

Patent metadata

Field	Value
Publication number	US-10351610-B2
Application number	US-201313757674-A
Country	US
Kind code	B2
Filing date	Feb 1, 2013
Priority date	Feb 2, 2012
Publication date	Jul 16, 2019
Grant date	Jul 16, 2019

Abstract

This disclosure concerns compositions and methods for increasing the expression of a polynucleotide of interest. Some embodiments concern novel transactivation polypeptides and variants thereof that have been identified in plants, and methods of using the same. Particular embodiments concern the use of at least one DNA-binding polypeptide in a fusion protein to target at least one transactivation polypeptide or variant thereof to a specific binding site on a nucleic acid comprising the polynucleotide of interest, such that its expression may be increased.

First claim

What may be claimed is:

1. A synthetic transcriptional activator fusion protein comprising:
a single DNA-binding peptide; and
a heterologous transactivation domain comprising the interaction motif peptide of SEQ ID NO:22.

2. The synthetic transcriptional activator fusion protein of claim 1 , wherein the DNA-binding peptide is selected from the group consisting of a zinc finger DNA-binding domain; a consensus binding sequence from a AVRBS3-inducible gene or synthetic binding sequence engineered therefrom; TAL; LexA; a Tet repressor; LacR; and a steroid hormone receptor.

3. The synthetic transcriptional activator fusion protein of claim 2 , wherein the DNA-binding peptide is a zinc finger DNA-binding domain.

4. The synthetic transcriptional activator fusion protein of claim 1 , wherein the transactivation domain comprises an amino acid sequence that is at least 80% identical to SEQ ID NO:107 or SEQ ID NO:108.

5. The synthetic transcriptional activator fusion protein of claim 1 , wherein the transactivation domain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO:107 or SEQ ID NO:108.

6. The synthetic transcriptional activator fusion protein of claim 1 , wherein the transactivation domain comprises SEQ ID NO:107 or SEQ ID NO:108.

7. A nucleic acid molecule comprising a polynucleotide encoding the synthetic transcriptional activator fusion protein of claim 1 , the polynucleotide comprising:
a first nucleotide sequence encoding the DNA-binding peptide; and
a second nucleotide sequence encoding the heterologous transactivation domain,
wherein the first and second nucleotide sequences are expressed from the nucleic acid molecule in frame and in a single transcript.

8. The nucleic acid molecule of claim 7 , wherein the first and second nucleotide sequences are separated by a third nucleotide sequence in the polynucleotide.

9. The nucleic acid molecule of claim 7 , wherein the polynucleotide is operably linked to a gene regulatory element.

10. A vector comprising the nucleic acid molecule of claim 9 .

11. The nucleic acid molecule of claim 9 , wherein the gene regulatory element is a promoter that is functional in a plant cell.

12. A host cell comprising the nucleic acid molecule of claim 9 .

13. The host cell of claim 12 , wherein the host cell is a plant cell or a yeast cell.

14. A plant cell comprising the nucleic acid molecule of claim 11 .

15. The plant cell of claim 14 , wherein the polynucleotide and gene regulatory element are integrated into the genome of the cell.

16. A transgenic plant material comprising the nucleic acid molecule of claim 11 .

17. The transgenic plant material of claim 16 , wherein the plant material is a plant cell, plant cell culture, plant tissue, plant tissue culture, plant part, plant commodity product, or whole plant.

18. The transgenic plant material of claim 17 , wherein the plant material is a plant cell, plant cell culture, plant tissue, plant tissue culture, plant part, or whole plant.

19. A method for increasing the expression of a polynucleotide of interest in a host cell, the method comprising:
introducing the nucleic acid molecule of claim 9 into a host cell comprising the polynucleotide of interest, wherein the polynucleotide of interest is operably linked to a second polynucleotide that binds specifically to the DNA-binding peptide, thereby increasing the expression of the polynucleotide of interest in the host cell.

20. The method according to claim 19 , wherein the host cell is a plant cell.

21. The method according to claim 20 , wherein introducing the nucleic acid molecule into the host cell comprises crossing a plant comprising the polynucleotide encoding the synthetic transcriptional activator fusion protein with a plant comprising the host cell.

22. The method according to claim 19 , wherein the introducing into the host cell a nucleic acid molecule comprises transforming said host cell with the nucleic acid molecule encoding the synthetic transcriptional activator protein.

23. The nucleic acid molecule of claim 7 , wherein the synthetic transcriptional activator fusion protein further comprising a promoter and a 3′UTR.

Assignees

• Dow Agrosciences Llc

Inventors

• Petolino Joseph
• Li Jianquan
• Evans Steven L
• Blue Ryan C

Classifications

• C12N15/8216 Primary

  Methods for controlling, regulating or enhancing expression of transgenes in plant cells · CPC title

• C12Y203/01183

  Phosphinothricin acetyltransferase (2.3.1.183) · CPC title

• C12N15/8217

  Gene switch · CPC title

• C07K2319/81

  containing a Zn-finger domain for DNA binding · CPC title

• C07K2319/71

  containing domain for transcriptional activaation, e.g. VP16 · CPC title