METHODS AND COMPOSITIONS FOR INACTIVATING ALPHA 1,6 FUCOSYLTRANSFERASE (FUT8) GENE EXPRESSION

摘要

Disclosed herein are methods and compositions for inactivating a FUT8 gene, using fusion proteins comprising a zinc finger protein and a cleavage domain or cleavage half-domain. Polynucleotides encoding said fusion proteins are also provided, as are cells comprising said polynucleotides and fusion proteins.

主权项

1. A method of producing an antibody in an isolated cell, the method comprising:
inactivating an endogenous cellular FUT8 gene in an isolated cell by introducing one or more nucleic acids encoding first and second zinc finger nucleases, each zinc finger nuclease comprising a cleavage domain and a zinc finger protein, wherein the at least one of the zinc finger nucleases comprises a zinc finger DNA-binding domain comprising four, five or six zinc finger recognition regions ordered F1 to F4, F1 to F5 or F1 to F6 from N-terminus to C-terminus, and wherein F1 to F4, F1 to F5 or F1 to F6 comprise amino acid sequences as shown in Table 1, wherein the first and second zinc finger nucleases cleave and inactivate the endogenous cellular FUT8 gene in the isolated cell; and introducing an exogenous sequence encoding an antibody into the cell, wherein the antibody is produced in the isolated cell.