Genetic studies have demonstrated that loss of the transcriptional co-activator p/CIP leads to resistance to obesity and diabetes, especially in extreme mouse obesity models. The goal of this research is to find small molecules that decrease p/CIP protein stability, thus providing new understanding and novel approaches to cure obesity and diabetes. In an in vitro white fat differentiation system, more ..
Target
Depositor Specified Assays
| AID | Name | Type | Comment |
|---|---|---|---|
| 1984 | Fluorescence for the identification of compounds that decrease p/CIP protein stability | confirmatory | uHTS fluorescence for the identification of compounds that decrease p/CIP protein stability |
| 2005 | uHTS fluorescence for the identification of compounds that decrease EGFP protein stability | confirmatory | EGFP counter screen |
Description:
Data Source: Sanford-Burnham Center for Chemical Genomics (SBCCG)
Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA)
Network: NIH Molecular Libraries Production Centers Network (MLPCN)
Grant Number: 1 R03 MH084852-01
Assay Provider: Zhiyong Wang PhD, The Salk Institute for biological studies, La Jolla CA
Genetic studies have demonstrated that loss of the transcriptional co-activator p/CIP leads to resistance to obesity and diabetes, especially in extreme mouse obesity models. The goal of this research is to find small molecules that decrease p/CIP protein stability, thus providing new understanding and novel approaches to cure obesity and diabetes. In an in vitro white fat differentiation system, p/CIP is important for fat cell formation. A fluorescence-based primary assay employing this in vitro cellular model has been developed to screen for small molecules that destabilize a p/CIP-EGFP fusion protein. Individual compounds will be incubated with the stably transfected cell line containing this GFP (green fluorescence) fusion protein. Small molecules will be identified that only diminish the GFP signal from the p/CIP-EGFP fusion protein cells.
The primary screen, AID 1984, is a cell-based HTS assay that utilizes an NIH3T3 derived subline F442A, a fibroblast line capable of differentiation into adipocytes in vitro, transfected with a p/CIP-EGFP fusion protein driven by a CMV promoter. Drugs interfering with p/CIP expression levels should directly affect the level of EGFP expression. The assay measures total fluorescence intensity of EGFP.
A counter assay, AID 2005, utilizes an NIH3T3 derived subline F442A, a fibroblast line capable of differentiation into adipocytes in vitro transfected with an EGFP protein driven by a CMV promoter. The assay measures total fluorescence intensity of EGFP and is used to identify false positive hits for compounds that target only EGFP for degradation.
No hits found in these assays were progressed to SAR analysis. Work has stopped on this project.
Source Affiliation: Sanford-Burnham Medical Research Institute (SBMRI, San Diego, CA)
Network: NIH Molecular Libraries Production Centers Network (MLPCN)
Grant Number: 1 R03 MH084852-01
Assay Provider: Zhiyong Wang PhD, The Salk Institute for biological studies, La Jolla CA
Genetic studies have demonstrated that loss of the transcriptional co-activator p/CIP leads to resistance to obesity and diabetes, especially in extreme mouse obesity models. The goal of this research is to find small molecules that decrease p/CIP protein stability, thus providing new understanding and novel approaches to cure obesity and diabetes. In an in vitro white fat differentiation system, p/CIP is important for fat cell formation. A fluorescence-based primary assay employing this in vitro cellular model has been developed to screen for small molecules that destabilize a p/CIP-EGFP fusion protein. Individual compounds will be incubated with the stably transfected cell line containing this GFP (green fluorescence) fusion protein. Small molecules will be identified that only diminish the GFP signal from the p/CIP-EGFP fusion protein cells.
The primary screen, AID 1984, is a cell-based HTS assay that utilizes an NIH3T3 derived subline F442A, a fibroblast line capable of differentiation into adipocytes in vitro, transfected with a p/CIP-EGFP fusion protein driven by a CMV promoter. Drugs interfering with p/CIP expression levels should directly affect the level of EGFP expression. The assay measures total fluorescence intensity of EGFP.
A counter assay, AID 2005, utilizes an NIH3T3 derived subline F442A, a fibroblast line capable of differentiation into adipocytes in vitro transfected with an EGFP protein driven by a CMV promoter. The assay measures total fluorescence intensity of EGFP and is used to identify false positive hits for compounds that target only EGFP for degradation.
No hits found in these assays were progressed to SAR analysis. Work has stopped on this project.
Protocol
Please see pertinent AIDs: 1984, 2005
Comment
A probe development effort based on the hits from the HTS screening campaign was unsuccessful. No probes were identified and the SBCCG probe development effort for this project has now stopped.
Additional Information
Grant Number: 1 R03 MH084852-01
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