Targeted integration into the PPP1R12C locus
First Claim
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1. A method for expressing at least one product of an exogenous nucleic acid sequence in a cell, the method comprising:
- (a) expressing a first fusion protein in the cell, the first fusion protein comprising a first DNA-binding domain and a first cleavage domain or cleavage half-domain, wherein the first DNA-binding domain has been engineered to bind to a first target site in the PPP1R12C gene in the genome of the cell; and
(b) contacting the cell with a polynucleotide comprising an exogenous nucleic acid sequence under conditions such that the first fusion protein cleaves the genome of the cell in the PPP1R12C gene and the exogenous nucleic acid sequence is inserted in a homology dependent manner into the cleavage site and expressed.
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Abstract
Disclosed herein are methods and compositions for targeted integration of an exogenous sequence into the human PPP1R12C locus, for example, for expression of a polypeptide of interest.
121 Citations
35 Claims
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1. A method for expressing at least one product of an exogenous nucleic acid sequence in a cell, the method comprising:
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(a) expressing a first fusion protein in the cell, the first fusion protein comprising a first DNA-binding domain and a first cleavage domain or cleavage half-domain, wherein the first DNA-binding domain has been engineered to bind to a first target site in the PPP1R12C gene in the genome of the cell; and (b) contacting the cell with a polynucleotide comprising an exogenous nucleic acid sequence under conditions such that the first fusion protein cleaves the genome of the cell in the PPP1R12C gene and the exogenous nucleic acid sequence is inserted in a homology dependent manner into the cleavage site and expressed. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27)
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- 28. An engineered zinc finger protein that binds to the PPP1R12C gene, wherein the protein comprises four zinc fingers and the amino acid sequence of the recognition region of each of the zinc fingers is as follows:
- 29. An engineered zinc finger protein that binds to the PPP1R12C gene, wherein the protein comprises four zinc fingers and the amino acid sequence of the recognition region of each of the zinc fingers is as follows:
Specification