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

• US 10,017,775 B2
• Filed: 03/28/2017
• Issued: 07/10/2018
• Est. Priority Date: 10/22/2009
• Status: Active Grant

• Current Assignee: Dow Agrosciences LLC (Corteva, Inc.), Sangamo Therapeutics, Inc.
• Original Assignee: Dow Agrosciences LLC (Corteva, Inc.), Sangamo Therapeutics, Inc.
• Inventors: DeKelver, Russell, Gupta, Manju, Miller, Jeffrey C., Novak, Stephen, Petolino, Joseph F.
• Primary Examiner(s): Keogh, Matthew R
• Application Number:
  US15/471,545
• Publication Number:
  US 20170204425A1
• Time in Patent Office: 469 Days
• Field of Search: None
• 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 plant comprising at least one plant cell in which a β

-ketoacyl-ACP synthetase (KAS) gene is mutated using a nuclease comprising a cleavage domain and a DNA-binding domain that binds to one of the target sites selected from the group consisting of SEQ ID NO;

24 to 27 present in the KAS gene, wherein the mutation modifies fatty acid content in the plant.

2. The plant of claim 1, wherein the mutation is an insertion and/or a deletion.

3. The plant of claim 1, wherein the mutation comprises a point mutation.

4. The plant of claim 2, wherein the insertion comprises insertion of a donor sequence.

5. The plant of claim 4, wherein the donor sequence comprises a coding sequence or an RNA product.

6. The plant of claim 5, wherein the coding sequence comprises a herbicide tolerance gene.

7. The plant of claim 1, wherein the nuclease is a zinc finger nuclease or TALEN.

8. The plant of claim 1, wherein the plant is a monocot.

9. The plant of claim 8, wherein the monocot is selected from the group consisting of maize, rice, barley, oats, wheat, sorghum, rye, sugarcane, pineapple, onion, banana, and coconut.

10. The plant of claim 1, wherein the plant is a dicot.

11. The plant of claim 10, wherein the dicot is selected from the group consisting of tobacco, tomato, sunflower, cotton, sugarbeet, potato, lettuce, melon, soybean, canola and alfalfa.

12. The plant of claim 1, wherein the plant is an oil-producing plant selected from the group consisting of Brassica, maize, soybean, crambe, mustard, castor bean, peanut, sesame, cotton, linseed, safflower, oil palm, flax, sunflower, and coconut.

13. The plant of claim 1, wherein the nuclease is introduced into the cell as a polynucleotide encoding the nuclease.

14. The plant of claim 1, wherein C16 fatty acid levels are increased and C18 fatty acid levels are decreased as is reduced as compared to a wild-type plant.

15. The seed or progeny of a plant according to claim wherein the seed or progeny comprises the mutated KAS gene.