Large-scale induction of protein-protein interactions to discover novel effector proteins for proximity-based therapeutics
AssigneeThe Children's Hospital of Philadelphia
InventorsGeorge BURSLEM, Stephanie SANSBURY, Yevgeniy SEREBRENIK, Ophir SHALEM
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Abstract
Provided herein are high throughput methods of identifying a target/effector protein pair, wherein the effector protein has a desired effect on the target protein following chemical induction of dimerization. Protein-protein interactions are induced between a target of interest and any other protein using a generic heterobifunctional ligand that binds to domains introduced by pooled protein tagging. These methods provide for the high-throughput discovery of endogenous effectors for any phenotype of interest.
Claims33
A method of identifying a target/effector protein pair, wherein the effector protein has a desired effect on the target protein following chemical induction of dimerization, the method comprising:
- (i) obtaining a population of cells in which the target protein comprises or is tagged with a ligand binding domain;
- (ii) endogenously tagging potential effector proteins with orthogonal ligand binding domains, such that each cell in the population of cells comprises a single effector protein tagged with an orthogonal ligand binding domain and an integrated molecular barcode;
- (iii) contacting the population of cells with a heterobifunctional molecule that binds to both the ligand binding domain and the orthogonal ligand binding domain, thereby inducing protein-protein interactions between the target protein and the potential effector proteins;
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- (iv) identifying the potential effector proteins that induce the desired effect on the target protein by selecting cells with a phenotype of interest.
Dependencies are inferred from the claim text.