US 11,987,804 B2
Rapamycin resistant cells
Andrew M. Scharenberg, Seattle, WA (US); David J. Rawlings, Seattle, WA (US); Karen Sommer, Seattle, WA (US); Samuel West, Seattle, WA (US); Yuchi Chiang Honaker, Seattle, WA (US); and Ryo Takeuchi, Seattle, WA (US)
Assigned to Seattle Children's Hospital, Seattle, WA (US)
Appl. No. 17/049,782
Filed by Seattle Children's Hospital, Seattle, WA (US)
PCT Filed Apr. 25, 2019, PCT No. PCT/US2019/029118
§ 371(c)(1), (2) Date Oct. 22, 2020,
PCT Pub. No. WO2019/210057, PCT Pub. Date Oct. 31, 2019.
Claims priority of provisional application 62/663,562, filed on Apr. 27, 2018.
Prior Publication US 2021/0340573 A1, Nov. 4, 2021
Int. Cl. C12N 15/90 (2006.01); C07K 14/715 (2006.01); C12N 15/113 (2010.01); C12N 15/86 (2006.01)
CPC C12N 15/90 (2013.01) [C07K 14/7155 (2013.01); C12N 15/113 (2013.01); C12N 15/86 (2013.01); C07K 2317/53 (2013.01); C07K 2319/02 (2013.01); C07K 2319/03 (2013.01); C12N 2740/16043 (2013.01); C12N 2750/14143 (2013.01)] 29 Claims
 
1. A system comprising:
a first polynucleotide comprising a nucleotide sequence encoding a soluble FK506-binding protein (FKBP)-rapamycin-binding (FRB) domain polypeptide, wherein the soluble FRB domain polypeptide comprises an amino acid sequence having at least 90% identity to SEQ ID NO: 1;
(ii) a second polynucleotide encoding a first chemically induced signaling complex (CISC) component, wherein the first CISC component comprises a first extracellular binding domain comprising an FKBP domain, a first transmembrane domain, and a first cytoplasmic signaling domain; and
(iii) a third polynucleotide encoding a second CISC component, wherein the second CISC component comprises a second extracellular binding domain comprising an FRB domain, a second transmembrane domain, and a second cytoplasmic signaling domain; and
wherein the first CISC component and the second CISC component dimerize in the presence of a ligand selected from rapamycin or a rapalog to create a signaling-competent CISC.