Engineered zinc finger proteins targeting plant genes involved in fatty acid biosynthesis

• US 20110167521A1
• Filed: 10/22/2010
• Published: 07/07/2011
• Est. Priority Date: 10/22/2009
• Status: Active Grant

• Current Assignee: Dow Agrosciences LLC (Corteva, Inc.), Sangamo Biosciences, Inc.
• Original Assignee: Dow Agrosciences LLC (Corteva, Inc.), Sangamo Biosciences, Inc.
• Inventors: DeKelver, Russell, Gupta, Manju, Miller, Jeffrey C., Petolino, Joseph F., Novak, Stephen
• Granted Patent: US 8,592,645 B2
• Time in Patent Office: Days
• US Class Current: 800/298
• CPC Class Codes: C07K 14/415 from plants
• CPC Class Codes: C12N 15/8216 Methods for controlling, regulating or enhancing expression of transgenes in plant cells
• CPC Class Codes: C12N 15/8218 Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]
• CPC Class Codes: C12N 15/8247 involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12N 9/22 Ribonucleases RNAses, DNAses catalytic nucleic acids C12N15/113
• CPC Class Codes: C12Y 203/01041 Beta-ketoacyl-acyl-carrier-protein synthase I (2.3.1.41)
• CPC Class Codes: C12Y 301/00 Hydrolases acting on ester bonds (3.1)

Abstract

The present disclosure relates to engineered zinc finger proteins that target genes in plants involved in fatty acid biosynthesis. Methods of using such zinc finger proteins in modulating gene expression, gene inactivation, and targeted gene modification are also provided.

Claims

1. A non-naturally occurring zinc finger protein that modulates expression of at least one endogenous plant gene, wherein the at least one plant gene is involved in fatty acid biosynthesis and is selected from the group consisting of a β

-ketoacyl-ACP synthetase (KAS) gene, a Fatty acid thioesterase B (FatB) gene, a Fatty acid synthase (FAS) gene, a Fatty acid elongase gene, a Fatty acid thioesterase A (FatA) gene, a plastidial G-3-P dehydrogenase (GPDH) gene, a glycerokinase (GK) gene, a stearoyl-acyl carrier protein desaturase (S-ACP-DES) gene, and an oleoyl-ACP hydrolase gene.

2. A fusion protein comprising a zinc finger protein of claim 1 and a functional domain.

3. The fusion protein of claim 2, wherein the functional domain is a transcriptional regulatory domain.

4. The fusion protein of claim 2, wherein the functional domain is a cleavage domain.

5. The zinc finger protein of claim 1, wherein the plant gene is contained within a Brassica plant.

6. The fusion protein of claim 2, wherein the plant gene is contained within a Brassica plant.

7. A polynucleotide encoding one or more zinc finger proteins of claim 1.

8. A method of modifying one or more genes involved in fatty acid biosynthesis in a plant cell, the method comprising:

introducing, into the plant cell, one or more expression vectors comprising at least one polynucleotide according to claim 7, under conditions such that the one or more zinc finger proteins are expressed and the one or more genes are modified.

9. The method of claim 8, wherein the modification comprises modulating expression of at least one of the genes.

10. The method of claim 9, wherein expression of at least one gene is activated.

11. The method of claim 9, wherein expression of at least one gene is repressed.

12. The method of claim 7, wherein the modification comprises cleaving at least one of the genes.

13. The method of claim 12, further comprising the step of introducing a donor nucleic acid such that the donor vector is introduced into the site of cleavage by homologous recombination.

14. A plant cell comprising at least one gene that has been modified by the method of claim 8.

15. The plant cell of claim 14, wherein the cell is within a seed and fatty acid content in the seed is modified.

16. A plant comprising at least one cell according to claim 14.

17. A plant comprising at least one cell according to claim 15.

18. The seed or progeny of a plant according to claim 14.

19. The seed or progeny of a plant according to claim 15.

20. A plant comprising at least one cell comprising at least one zinc finger protein according to claim 1.

21. The seed or progeny of a plant according to claim 20.

22. A plant comprising at least one cell comprising at least one polynucleotide according to claim 7.

23. The seed or progeny of a plant according to claim 22.