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

• US 9,631,201 B2
• Filed: 11/25/2013
• Issued: 04/25/2017
• 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., Novak, Stephen, Petolino, Joseph F.
• Primary Examiner(s): KEOGH, MATTHEW R
• Application Number:
  US14/089,260
• Publication Number:
  US 20140157458A1
• Time in Patent Office: 1,247 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 non-naturally occurring zinc finger protein that modulates expression of at least one endogenous Fatty acid thioesterase B (FatB) plant gene, wherein the zinc finger proteins binds to one of the target sites selected from the group consisting of SEQ ID NO:

47-52.

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 zinc finger protein of claim 1, wherein the FatB gene is a Brassica gene.

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

6. The zinc finger protein of claim 4, wherein the zinc finger protein modulates expression of a FatB4 or FatB5 gene.

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

8. A method of modifying expression of 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 modify the expression of the one or more Fat genes.

9. The method of claim 8, wherein the modification comprises modulating expression of at least one FatB gene is modulated.

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

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

12. The method of claim 8, wherein the polynucleotide encodes a zinc finger nuclease and at least one FatB gene is cleaved.

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

14. A plant comprising at least one cell comprising the zinc finger protein of claim 1.

15. A plant comprising at least one cell comprising the polynucleotide of claim 7.