Methods and compositions for inactivating glutamine synthetase gene expression
First Claim
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1. A zinc finger DNA-binding domain comprising four, five or six zinc finger recognition regions as shown in Table 1.
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Abstract
Disclosed herein are methods and compositions for inactivating a glutamine synthetase (GS) 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.
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Citations
18 Claims
- 1. A zinc finger DNA-binding domain comprising four, five or six zinc finger recognition regions as shown in Table 1.
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7. A cell line in which glutamine synthetase (GS) is partially or fully inactivated by a nuclease.
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8. A method of inactivating an endogenous cellular GS gene in a cell, the method comprising:
(a) introducing, into a cell, a first nucleic acid encoding a first polypeptide, wherein the first polypeptide comprises; (i) a zinc finger DNA-binding domain that is engineered to bind to a first target site in an endogenous GS gene; and (ii) a cleavage domain;
such that the polypeptide is expressed in the cell, whereby the polypeptide binds to the target site and cleaves the GS gene.- View Dependent Claims (9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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