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US 12,595,492 B2Granted
Rapamycin resistant cells
AssigneeSeattle Children's Hospital
InventorsAndrew M. Scharenberg, David J. Rawlings, Karen Sommer, Samuel West, Yuchi Chiang Honaker, Ryo Takeuchi
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Abstract
The present application relates to, inter alia, compositions including proteins for expression in host cells to render them resistant to rapamycin. The application further relates to methods of using the proteins, cells, and compositions disclosed therein for modulating cell signaling and for selective expansion of cells.
Claims32
A cell prepared by introducing into a cell:
- (i) a first polynucleotide encoding a soluble FK506-binding protein (FKBP)-rapamycin-binding (FRB) domain polypeptide;
- (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 to create a signaling-competent CISC.
3 more elements
A cell comprising:
- (i) a first polynucleotide encoding a soluble FK506-binding protein (FKBP)-rapamycin-binding (FRB) domain polypeptide;
- or the soluble FRB domain polypeptide;
- (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;
- or the first CISC component;
- 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;
- or the second CISC component;
- and
- wherein the first CISC component and the second CISC component dimerize in the presence of a ligand to create a signaling-competent CISC.
6 more elements
A method for preparing a cell, comprising introducing into a cell:
- (i) a first polynucleotide encoding a soluble FK506-binding protein (FKBP)-rapamycin-binding (FRB) domain polypeptide or the soluble FRB domain polypeptide;
- (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 to create a signaling-competent CISC.
3 more elements
Dependencies are inferred from the claim text.