Patent ID: 11884930
Assignee: CITY OF HOPE
Field: Measurement (Instruments)
Classification: CPC C  A  G | IPC C  G

Claim 0:
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.