Novel screening system for drug target antagonist based on cell life/death phenotype
OwnerHai JIANG, Shishuang CHEN +1inventors
InventorsHai JIANG, Shishuang CHEN, Yankai CHU
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Abstract
Provided is a novel screening system for a drug target antagonist based on cell life/death phenotype. Specifically, on the basis of a target protein and a conditional suicide protein FKBP 12 (F36V)-Caspase9, a screening system is designed for screening a target protein inhibitor or a degradation agent. The screening system is stable and efficient, the phenotype is obviously easy to observe, and the screening system can be used for high-throughput screening.
Claims17
A screening system for screening inhibitors against target proteins, wherein, the screening system comprises a culture system and the following components present in the culture system:
- (a) a live detection cell, which expresses an exogenous fusion protein that is a fusion protein of a target protein and a cell suicide element, wherein the fusion protein induces apoptosis of the detection cell in the presence of a suicide inducer;
- (b) a test object to be screened;
- wherein, when the detection cell expresses the fusion protein and the test object does not cause degradation or reduction of the fusion protein, the suicide inducer induces apoptosis of the detection cell;
- and,
- when the test object causes the detection cell not to express the fusion protein or cause degradation or reduction of the fusion protein, the suicide inducer does not induce apoptosis of the detection cell.
A method for screening candidate drugs for a target protein, comprising the steps of:
- (a) taking a culture system added with a test object to be screened as an experimental group;
- and taking a culture system without adding a test compound as a blank control group, wherein the culture system contains a cultured live detection cell expressing a fusion protein of an exogenous, the fusion protein being a fusion protein of a target protein and a cell suicide element, wherein the fusion protein induces apoptosis of the detection cell in the presence of a suicide inducer;
- and
- (b) adding a suicide inducer to the experimental group and blank control group, and observing the survival of cells detected in the experimental group and blank control group;
- wherein, when the survival number of cells detected in the experimental group is significantly higher than that of the control group, it is indicated that the test object is a candidate drug.
A fusion protein, wherein the structure of the fusion protein is as shown in the Formula I:
- B-L1-A Formula I
- wherein,
- B is a disease target protein;
- A is a cell suicide element;
- L1 is none, or linker sequence;
- “-” is independently a linking peptide or a peptide bond.
Dependencies are inferred from the claim text.