Universal donor system for gene targeting
View Patent ↗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.
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.