Chimeric polypeptides and methods of altering the membrane localization of the same
AssigneeCell Design Labs, Inc.
InventorsStephen Santoro, Scott Coyle, Levi Rupp, Peter Emtage
Life of this patent
Estimating the term…
Abstract
Methods and compositions are provided for reversibly localizing proteins to the exterior part of the cell surface. Compositions provided herein can include nucleic acids that encode polypeptides of interest and the ligand binding domain (LBD) of a nuclear hormone receptor. Medical applications are provided, including controlling the toxicity of CAR T cells.
Claims18
A method of inducing membrane localization of at least a portion of a single-chain chimeric polypeptide to an extracellular side of the membrane in a mammalian cell, the method comprising:
- contacting a mammalian cell expressing a single-chain chimeric polypeptide with an amount of amoxifen sufficient to induce localization of at least a portion of the single-chain chimeric polypeptide to an extracellular side of the membrane of the cell;
- wherein the single-chain chimeric polypeptide comprises the amino sequences of SEQ ID NO:
- 31, SEQ ID NO:
- 33 or SEQ ID NO:
- 35.
The method of 9, wherein the contacting step results in the treatment of the blood cancer in the mammal.
A method of reversibly altering membrane localization of a single-chain chimeric polypeptide in a mammalian cell, the method comprising:
- (a) contacting a mammalian cell expressing a single-chain chimeric polypeptide with an amount of amoxifen sufficient to induce localization of at least a portion of the single-chain chimeric polypeptide to an extracellular side of the plasma membrane of the cell;
- and
- (b) contacting the mammalian cell with a reduced amount of amoxifen that results in a decreased level of the single-chain chimeric polypeptide on an extracellular side of the membrane of the mammalian cell;
- wherein the single-chain chimeric polypeptide comprises the amino sequences of SEQ ID NO:
- 31, SEQ ID NO:
- 33 or SEQ ID NO:
- 35.
A method of inducing plasma membrane localization of at least a portion of a single-chain chimeric polypeptide to an extracellular side of the plasma membrane in a mammalian cell, the method comprising:
- contacting a mammalian cell expressing a single-chain chimeric polypeptide with an amount of tamoxifen sufficient to induce localization of at least a portion of the single-chain chimeric polypeptide to an extracellular side of the plasma membrane of the cell;
- wherein the single-chain chimeric polypeptide comprises the amino acid sequence of SEQ ID NO:
- 31, SEQ ID NO:
- 33 or SEQ ID NO:
- 35.
Dependencies are inferred from the claim text.