Patent ID: 9249428
Filing Date: 2016-02-02
CPC Classification: C12N

Claim Text:
1. A method for creating a targeted deletion of specific length and specific borders in a region of interest in a genomic locus in a host cell, the method comprising introducing first and second nuclease pairs and a donor polynucleotide into the host cell, wherein (i) the first and second nuclease pairs each comprise a left and a right nuclease, each nuclease comprising DNA-binding domain that recognizes a target sequence in the region of interest of the genomic locus, (ii) the first and second nuclease pairs cleave in the region of interest genomic locus; (iii) the left nuclease of each nuclease pair recognizes a target sequence 5′ to the right nuclease; (iv) the first nuclease pair binds to target sequences 3′ to the target sequences bound by the second nuclease pair; (v) the donor polynucleotide comprises the target sequences for the left nuclease of the second nuclease pair and for the right nuclease of the first nuclease pair and regions of homology to the genomic locus and further wherein the donor polynucleotide includes a deletion relative to the target sequence of specific length and specific borders, wherein the deletion extends between the target sequence for the left nuclease of the second nuclease pair and the target sequence for the right nuclease of the first nuclease pair; and (vi) the donor polynucleotide is introduced into the genomic locus such that a targeted deletion of specific length and specific borders is generated in the genomic locus.