Modified FRT recombination site libraries and methods of use

• US 9,777,284 B2
• Filed: 12/18/2015
• Issued: 10/03/2017
• Est. Priority Date: 07/18/2005
• Status: Active Grant

• Current Assignee: Pioneer Hi-Bred International (Corteva, Inc.)
• Original Assignee: Pioneer Hi-Bred International (Corteva, Inc.)
• Inventors: Tao, Yumin, Bidney, Dennis L., Gordon-Kamm, William J., Lyznik, Leszek A.
• Primary Examiner(s): Page, Brent
• Application Number:
  US14/974,806
• Publication Number:
  US 20160108412A1
• Time in Patent Office: 655 Days
• Field of Search: None
• CPC Class Codes: C12N 15/102 Mutagenizing nucleic acids
• CPC Class Codes: C12N 15/1051 Gene trapping, e.g. exon-, intron-, IRES-, signal sequence-trap cloning, trap vectors
• CPC Class Codes: C12N 15/1093 General methods of preparing gene libraries, not provided for in other subgroups
• CPC Class Codes: C12N 15/8213 Targeted insertion of genes into the plant genome by homologous recombination
• CPC Class Codes: C40B 40/08 Libraries containing RNA or DNA which encodes proteins, e.g. gene libraries

Abstract

Methods and compositions using populations of randomized modified FRT recombination sites to identify, isolate and/or characterize modified FRT recombination sites are provided. Kits comprising the library populations of FRT sites are also provided, as are methods to make a library of modified FRT recombination sites. The recombinogenic modified FRT recombination sites can be employed in a variety of methods for targeted recombination of polynucleotides of interest.

Claims

1. A plant cell having stably incorporated into its genome at least a first nucleotide sequence of interest flanked by a first and a second recombination site, wherein said first and said second recombination sites are dissimilar and non-recombinogenic with respect to one another;

such that the genome of the plant cell comprises in the following order the first recombination site, the first nucleotide sequence of interest, and the second recombination site, wherein said first and said second recombination sites are selected from the group consisting of FRT1 (SEQ ID NO;

39) and FRT87 (SEQ ID NO;

24).

2. The plant cell of claim 1, wherein said plant cell is from a monocot.

3. The plant cell of claim 2, wherein said monocot is maize or wheat.

4. The plant cell of claim 1, wherein said plant cell is from a dicot.

5. The plant cell of claim 4, wherein said dicot is canola, Brassica, soybean, sunflower, or cotton.

6. The plant cell of claim 1, wherein said plant cell further comprises a LOX site or a biologically active variant of the LOX site stably incorporated into its genome.

7. A plant comprising the plant cell of claim 1.

8. A transformed seed comprising the plant cell of claim 1.