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Genome editing of a;locus using nucleases

  • US 9,567,573 B2
  • Filed: 04/25/2011
  • Issued: 02/14/2017
  • Est. Priority Date: 04/26/2010
  • Status: Active Grant
First Claim
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1. An isolated fusion protein comprising a Fokl nuclease domain and one or more TAL effector (TALE) polypeptides comprising a plurality of TALE repeat domains, wherein each TALE repeat domain comprises a RVD, wherein:

  • (i) each RVD of said TALE repeat domains bind to a target site in a Rosa gene, wherein said target site is selected from the group consisting of the nucleotide sequence of SEQ ID NO;

    28, SEQ ID NO;

    29, SEQ ID NO;

    79, SEQ ID NO;

    80, SEQ ID NO;

    81, SEQ ID NO;

    82, SEQ ID NO;

    87, SEQ ID NO;

    88, SEQ ID NO;

    89, and SEQ ID NO;

    90; and

    further wherein said RVD comprises the amino acids NI, HI, Al, CI, DI, El, RI or KI which recognizes adenine (A) in said target site;

    the amino acids HD, AD, KD, YD, ED, AD, ND or RD which recognizes cytosine (C) in said target site;

    the amino acids NN, DK, DH, KN, EN, AN, CN, GN, FN, AK, NK or CK which recognizes guanine (G) in said target site; and

    the amino acids NG, KG MG, QG, RG, AA, QA, IG, IP, LA, YG, HG, SG, VG, IA or VA which recognizes thymine (T) in said target site; and

    (ii) said fusion protein cleaves said Rosa gene.

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