FAD2 FUNCTIONAL LOCKS AND RELATED SPECIFIC FOR TARGET WEBSITE CONNECTING PROTEINS ABLE TO INDUCE DIRECTED GAP

摘要

1. A method for modifying the FAD2 gene in a soy cell, comprising: splitting a site-specific image of a target site in the FAD2 gene of a soy cell to create a gap in the FAD2 gene, wherein the FAD2 gene is modified after cleavage. 2. The method of claim 1, further comprising integrating into the gap the nucleotide sequence of interest. The method of claim 1 or 2, wherein the FAD2 gene is a FAD2 2.3, FAD2 2.6 gene, or both. The method of claim 1 or 2, wherein the site-specific cleavage comprises introducing a fusion protein containing a DNA binding domain and a cleaving domain or a cleaving half-domain into a cell, or a polynucleotide encoding a fusion protein, wherein the fusion protein specifically binds to the site is a target and carries out cleavage at, or near the target site to create, thus, a gap. The method of claim 4, wherein the DNA binding domain is selected from the group consisting of the DNA binding domain of meganuclease, the DNA binding domain of leucine "zippers", similar to the transcription activator (TAL) of the DNA binding domain, RNA-directed CRISPR-Cas9 , recombinases, the DNA-binding domain of the protein with zinc fingers, as well as chimeric combinations of any of the above. 6. The method of claim 4, wherein the cleavage domain or cleavage half-domain is selected from the group consisting of cleaving half-domain of a restriction endonuclease type IIS and a homing endonuclease. The method of claim 4, wherein the fusion protein is a zinc finger nuclease. The method of claim 7, wherein the zinc finger nuclease comprises three to six zinc finger domains, each zinc finger domain containing a helical recognition region, and the zinc finger protein containing helical regions