US 10,888,589 B2
Modulation of stem cells using zinc finger proteins
Alan P. Wolffe, Richmond, CA (US); Michael Moore, London (GB); Timothy Farries, Middlesex (GB); Trevor Collingwood, Richmond, CA (US); and Fyodor Urnov, Richmond, CA (US)
Assigned to Sangamo Therapeutics, Inc., Richmond, CA (US)
Filed by Sangamo Therapeutics, Inc., Richmond, CA (US); and Elizabeth J. Wolffe, Richmond, CA (US)
Filed on Apr. 13, 2017, as Appl. No. 15/486,731.
Application 14/270,038 is a division of application No. 10/490,787, granted, now 8,735,153, issued on May 27, 2014, previously published as PCT/US02/30413, filed on Sep. 24, 2002.
Application 15/486,731 is a continuation of application No. 14/270,038, filed on May 5, 2014, granted, now 9,624,509.
Claims priority of provisional application 60/324,619, filed on Sep. 24, 2001.
Claims priority of provisional application 60/367,252, filed on Mar. 21, 2002.
Claims priority of provisional application 60/374,176, filed on Apr. 17, 2002.
Prior Publication US 2017/0224741 A1, Aug. 10, 2017
Int. Cl. C12N 15/63 (2006.01); A61K 48/00 (2006.01); C07H 21/04 (2006.01); A61K 35/545 (2015.01); C12N 5/0735 (2010.01); G01N 33/50 (2006.01); C12N 15/85 (2006.01); C07K 14/74 (2006.01); C12N 15/62 (2006.01); A61K 35/12 (2015.01)
CPC A61K 35/545 (2013.01) [C07K 14/70539 (2013.01); C12N 5/0606 (2013.01); C12N 15/63 (2013.01); C12N 15/85 (2013.01); G01N 33/5008 (2013.01); G01N 33/5023 (2013.01); G01N 33/5073 (2013.01); A61K 35/12 (2013.01); C07H 21/04 (2013.01); C12N 15/62 (2013.01); C12N 2501/40 (2013.01); C12N 2503/02 (2013.01); C12N 2510/00 (2013.01)] 6 Claims
 
1. An isolated stem cell comprising a homeobox protein B4 (HOXB4) gene or octamer-binding transcription factor 4 (OCT-4) gene, wherein expression of the HOXB4 or OCT-4-gene is modified by an artificial fusion protein comprising
(i) an engineered C2H2 zinc finger protein (ZFP) DNA binding domain that binds to a target site in the HOXB4 or OCT-4 gene, the ZFP comprising a ZFP comprising 3 zinc finger domains ordered F1 to F3, each zinc finger domain comprising a recognition helix region as follows:
F1: RSDHLAR (SEQ ID NO:21);
F2: RSDELQR (SEQ ID NO:22);
F3: RSDERKR (SEQ ID NO:23)
for binding to the HOXB4 gene or a ZFP comprising 3 zinc finger domains ordered F1 to F3, each zinc finger domain comprising a recognition helix region as follows:
F1: RSDHLAR (SEQ ID NO:2);
F2: TSGSLTR (SEQ ID NO:3); and
F3: RSDNLAR (SEQ ID NO:4) for binding to OCT-4; and
(ii) a functional domain comprising a VP16 or p65 transcriptional activation domain or a KRAB repression domain.