US 11,884,930 B2
Method of inactivating a glucocorticoid receptor gene in an isolated cell
Andreas Reik, Brisbane, CA (US); Michael Jensen, Sierra Madre, CA (US); Michael C. Holmes, Brisbane, CA (US); Philip D. Gregory, Brisbane, CA (US); and Dale Ando, Brisbane, CA (US)
Assigned to Sangamo Biosciences, Inc., Brisbane, CA (US); and City of Hope, California (CA)
Filed by Sangamo Therapeutics, Inc., Brisbane, CA (US); and City of Hope, Duarte, CA (US)
Filed on Dec. 18, 2020, as Appl. No. 17/127,210.
Application 17/127,210 is a division of application No. 14/947,476, filed on Nov. 20, 2015, granted, now 10,907,175.
Application 14/947,476 is a continuation of application No. 11/983,888, filed on Nov. 13, 2007, granted, now 9,217,026, issued on Dec. 22, 2015.
Claims priority of provisional application 60/967,820, filed on Sep. 7, 2007.
Claims priority of provisional application 60/859,417, filed on Nov. 15, 2006.
Claims priority of provisional application 60/859,025, filed on Nov. 13, 2006.
Prior Publication US 2021/0102222 A1, Apr. 8, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 5/00 (2006.01); C12N 15/86 (2006.01); C12N 15/62 (2006.01); C07K 14/72 (2006.01); C12N 9/22 (2006.01); G01N 33/74 (2006.01)
CPC C12N 15/86 (2013.01) [C07K 14/721 (2013.01); C12N 9/22 (2013.01); C12N 15/62 (2013.01); G01N 33/743 (2013.01); C12N 2710/10043 (2013.01); C12N 2740/15043 (2013.01); G01N 2333/723 (2013.01)] 12 Claims
 
1. A method of selecting isolated human cells comprising a transgene integrated into an endogenous glucocorticoid receptor (GR) gene, the method comprising:
a) introducing:
i) one or more polynucleotides encoding a first and second pair of zinc finger nuclease (ZFN); and
ii) a polynucleotide comprising a transgene flanked by homology arms that target a human glucocorticoid receptor (GR) gene into isolated human cells such that the transgene is integrated into an endogenous GR gene,
wherein
the first ZFN comprises:
a zinc finger DNA-binding domain comprising the following recognition helices in the following order:
 
 (SEQ ID NO: 23)
 
 TSRALTA;
 
 
 
 (SEQ ID NO: 17)
 
 DRANLSR;
 
 
 
 (SEQ ID NO: 24)
 
 RSDNLSE;
 
 and
 
 
 
 (SEQ ID NO: 26)
 
 ERANRNS;
and
the second ZFN comprises:
a zinc finger DNA-binding domain comprising the following recognition helices in the following order:
 
 (SEQ ID NO: 33)
 
 DSWNLQV;
 
 
 
 (SEQ ID NO: 27)
 
 RSANLTR;
 
 
 
 (SEQ ID NO: 30)
 
 TSGNLTR;
 
 and
 
 
 
 (SEQ ID NO: 31)
 
 TSGSLTR;
or
the first ZFN comprises:
a zinc finger DNA-binding domain comprising the following recognition helices in the following order:
 
 (SEQ ID NO: 42)
 
 RSANLAR;
 
 
 
 (SEQ ID NO: 43)
 
 RSDNLRE;
 
 
 
 (SEQ ID NO: 44)
 
 QSSNLAR;
 
 and
 
 
 
 (SEQ ID NO: 45)
 
 QSADRTK;
and
the second ZFN comprises:
a zinc finger DNA-binding domain comprising the following recognition helices in the following order:
 
 (SEQ ID NO: 40)
 
 RQDCLSL;
 
 
 
 (SEQ ID NO: 37)
 
 RNDNRKT;
 
 
 
 (SEQ ID NO: 38)
 
 RSDNLSR;
 
 and
 
 
 
 (SEQ ID NO: 39)
 
 TNQNRIT;
and
b) contacting the isolated human cells obtained in step a) with a corticosteroid such that cells not comprising the integrated transgene are killed and cells comprising the transgene integrated into the endogenous GR gene are selected.