Universal donor system for gene targeting

Patent metadata

Field	Value
Publication number	US-10077449-B2
Application number	US-201414531710-A
Country	US
Kind code	B2
Filing date	Nov 3, 2014
Priority date	Nov 4, 2013
Publication date	Sep 18, 2018
Grant date	Sep 18, 2018

Abstract

A universal donor polynucleotide is described that can be inserted at targeted locations in plant genomes to facilitate rapid and high throughput integration of a donor molecule within a specific genomic location.

First claim

What is claimed is:

1. A polynucleotide donor cassette comprising a site specific nuclease binding domain, an analytical domain, and a plasmid domain, wherein said analytical domain comprises a polynucleotide sequence with at least 95% sequence identity to SEQ ID NO: 142.

2. A transgenic cell comprising the polynucleotide donor cassette of claim 1 .

3. The transgenic cell of claim 2 , wherein the cell is a transgenic plant cell.

4. A transgenic plant comprising the transgenic plant cell of claim 3 .

5. The polynucleotide donor cassette of claim 1 , wherein the polynucleotide donor cassette having the sequence of SEQ ID NO: 132.

6. The polynucleotide donor cassette of claim 1 , wherein the site specific nuclease binding domain comprises two or more site specific nuclease binding sequences.

7. The polynucleotide donor cassette of claim 1 , wherein the site specific nuclease binding domain is a zinc finger binding domain.

8. The polynucleotide donor cassette of claim 1 , wherein the analytical domain is a polynucleotide sequence consisting of SEQ ID NO: 142.

9. The polynucleotide donor cassette of claim 1 , wherein the [−] polynucleotide donor cassette having the sequence of SEQ ID NO: 133.

10. The polynucleotide donor cassette of claim 1 , wherein the polynucleotide donor cassette further comprises one or more homology arm sequences.

11. The polynucleotide donor cassette of claim 1 , wherein the analytical domain does not encode a peptide.

12. The polynucleotide donor cassette of claim 1 , wherein the analytical domain further comprises coding sequences that encode a peptide.

13. The polynucleotide donor cassette of claim 12 , wherein the analytical domain comprises a gene expression cassette comprising a transgene.

14. The polynucleotide donor cassette of claim 13 , wherein the transgene comprises a reporter gene.

15. The polynucleotide donor cassette of claim 14 , wherein the reporter gene is selected from the group consisting of a yfp gene, a gus gene, a rfp gene, a gfp gene, a kanamycin resistance gene, an aad-1 gene, an aad-12 gene, a pat gene, and a glyphosate tolerant gene.

Assignees

• Dow Agrosciences Llc

Inventors

• Sastry-Dent Lakshmi
• Evans Steven L
• Blue Ryan C
• Cao Zehui

Classifications

• C12N15/8209 Primary

  Selection, visualisation of transformants, reporter constructs, e.g. antibiotic resistance markers · CPC title

• C12N15/8212

  Colour markers, e.g. beta-glucoronidase [GUS], green fluorescent protein [GFP], carotenoid · CPC title

• C12N15/8213 Primary

  Targeted insertion of genes into the plant genome by homologous recombination · CPC title

• C12N15/8275

  Glyphosate · CPC title