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US 11,634,721 B2Granted
Reconstruction of site specific nuclease binding sites
AssigneeDow AgroSciences LLC
InventorsDavid R. Corbin, Wei Chen, Stephen Novak, Ryan M. Lee, Sandeep Kumar, Andrew Asberry, Andrew F. Worden
Life of this patent
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Abstract
Disclosed herein are methods and compositions for the repair of site specific nuclease binding sites by targeted integration and/or targeted excision of one or more sequences into a cell.
Claims15
A method for producing a repaired site specific nuclease binding site within a plant, the method comprising the following steps:
- a. Providing a plant genome comprising a first copy of a site specific nuclease binding site, an intervening polynucleotide sequence, and a second copy of the site specific nuclease binding site, wherein the first copy of the site specific nuclease binding site and the second copy of the site specific nuclease binding site are identical, and the first copy of the site specific nuclease binding site is located within 3,000 bp to 4,000 bp of the second copy of the site specific nuclease binding site;
- b. Introducing a site specific nuclease designed to bind and cleave at the site specific nuclease binding site;
- c. Cleaving the first copy of the site specific nuclease binding site;
- d. Cleaving the second copy of the site specific nuclease binding site;
- e. Excising the intervening polynucleotide sequence from the plant genome;
- f. Recombining the first cleaved site specific nuclease binding site with the second cleaved site specific nuclease binding site;
- g. Producing the repaired site specific nuclease binding site, wherein the repaired site specific nuclease binding site is repaired via a NHEJ-mediated cellular process and the repaired site specific nuclease binding site is identical to the first copy of the site specific nuclease binding site.
4 more elements
Dependencies are inferred from the claim text.