Target-related Tumors Research

1. Sample and: tissue, blood, cell sample acquisition;

2. Screening and verification of targets and related signal pathways:

1) Target screening: various chips, second-generation sequencing, etc.

2) Signal pathway: correlation analysis, such as lncRNA, microRNA detection, etc .;

3) Verification: in vitro verification and in vivo verification;

3. Functional omics verification of candidate targets and downstream pathway screening:

1) Cell level For specific targets, gain of function / loss of function is used to clarify the function of the target and the relationship with the tumor at the cell level;

2) Animal level Related research through KI / KO animal models;

4. Study on the signal pathways and action mechanisms of cell-level targets:

1) Through high-throughput screening (chips, proteomics, pull-down, yeast hybrids, literature, etc.) to further identify target upstream and downstream promoters, miRNA, LncRNA, and autophagy related genes;

2) Through gain of function / loss of function, verify the clear upstream and downstream relationship between the target and related molecular pathways at the cell level;

5.Signal pathways and action mechanisms of animal-level targets:

1) Prove a clear upstream and downstream relationship between the target and related molecular pathways through specific signaling pathway agonists / inhibitors

2) Further prove the relationship between the target and related molecular pathways through the gene gain of function / loss of function, and clarify the upstream and downstream relationships (Tg / KO mouse);

6. Detection of fine regulatory relationship between target and upstream and downstream target molecules:

1) Interaction between proteins: through Co-IP, pull-down, FRET;

2) Upstream and downstream transcriptional regulation: through promoter-luciferase system, EMSA, etc.

3) For more elaborate molecular interaction areas: study the interaction domains through mutations and truncated bodies, such as the search for fine domains of protein-protein interactions.

7. Establishment of drug screening model and drug screening:

Identify the molecular mechanisms of tumor targets and their related pathways, and establish a specific tumor drug screening model based on the characteristics of the target and its downstream signaling pathways, and use the high-throughput screening platform (HTS) to perform specific drugs from the compound library Filter.