Alternative splicing of pre-mRNAs plays a major role in the regulation of eukaryotic gene expression. Alternative splicing is now thought to be one of the primary reasons for the complexity of higher organisms leading to an average of three protein isoforms per gene. Aberrant splicing can lead to a number of diseases and is known to play a role in rare genetic diseases such as progeria syndrome. more ..
Related BioAssays
Related BioAssays
Targets
| Sequence: CDC-like kinase 4 [Homo sapiens] Description ..   Protein Family: Protein Kinases, catalytic domain Gene:CLK4 Related Protein 3D Structures |
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BioActive Compounds: 28
Depositor Specified Assays
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| AID | Name | Type | Probe | Comment |
|---|---|---|---|---|
| 1459 | Validation of Assay for Modulators of Lamin A Splicing | confirmatory | ||
| 1487 | qHTS Assay for Modulators of Lamin A Splicing | confirmatory | ||
| 1498 | Confirmation Concentration-Response Assay for Modulators of Lamin A Splicing | confirmatory | ||
| 1770 | qHTS Assay for Inhibitors of CDC-like Kinase 4 (Kinase-Glo Assay) | confirmatory | ||
| 1970 | Confirmation Assay for Inhibitors of CDC-like Kinase 4 (Kinase-Glo Assay) | confirmatory | ||
| 1997 | qHTS for Inhibitors of CDC-like Kinase 4: Summary | summary | 3 | |
| 488887 | qHTS for Inhibitors of CDC-like Kinase 1 and 4: Summary | summary | 1 | |
| 488872 | qHTS for Inhibitors of DYRK1A: Summary | summary | 1 | |
| 493204 | Confirmation Assay for Inhibitors of CDC-like Kinase 4 (Kinase-Glo Assay): SAR round 2 | confirmatory | ||
| 488883 | qHTS for Inhibitors of DYRK1B: Summary | summary | 1 |
Description:
NIH Molecular Libraries Probe Production Centers Network [MLPCN]
NIH Chemical Genomics Center [NCGC]
MLSCN Grant: 1R03MH084827-01
Assay Submitter (PI): Tom Misteli
NCGC Assay Overview:
Alternative splicing of pre-mRNAs plays a major role in the regulation of eukaryotic gene expression. Alternative splicing is now thought to be one of the primary reasons for the complexity of higher organisms leading to an average of three protein isoforms per gene. Aberrant splicing can lead to a number of diseases and is known to play a role in rare genetic diseases such as progeria syndrome. Hutchinson-Gilford Progeria Syndrome (HGPS) is a pediatric premature aging disease caused by a spontaneous mutation in the lamin A/C (LMNA) gene due to a single point mutation in the lamin A gene effecting 1 in 4 million people.
To identify both specific splicing modulators of the lamin A gene as well as general splicing modulators, a cell-based HTS assay was developed using a minigene reporter system (AIDs, 1459 & 1487). A quinazoline (CID: 16759567) showed weak activity as an inhibitor of aberrant splicing using the minigene reporter system (AID 1498). A preliminary kinase profile of this compound showed activity in splicing related kinases belonging to the CDC-like kinase (Clk) family particularly the Clk4. The Clk family is part of the CMGC group of protein kinases which includes Dyrks and are essential kinases found in all eukaryotes. Clks have been implicated to play a role in the regulation of alternative splicing [1]; while Dyrks, in particular over expression of Dyrk1A, has been linked to diseases as Down syndrome [2] and Alzheimer disease [3]. This AID summarizes the characterization of lead Clk4 inhibitors in a panel of these related kinases: Clk1, 2 & 3, DYRK1/DYRK1A, and DYRK1B. Potency against Clk4 was also confirmed in the panel.
NIH Chemical Genomics Center [NCGC]
MLSCN Grant: 1R03MH084827-01
Assay Submitter (PI): Tom Misteli
NCGC Assay Overview:
Alternative splicing of pre-mRNAs plays a major role in the regulation of eukaryotic gene expression. Alternative splicing is now thought to be one of the primary reasons for the complexity of higher organisms leading to an average of three protein isoforms per gene. Aberrant splicing can lead to a number of diseases and is known to play a role in rare genetic diseases such as progeria syndrome. Hutchinson-Gilford Progeria Syndrome (HGPS) is a pediatric premature aging disease caused by a spontaneous mutation in the lamin A/C (LMNA) gene due to a single point mutation in the lamin A gene effecting 1 in 4 million people.
To identify both specific splicing modulators of the lamin A gene as well as general splicing modulators, a cell-based HTS assay was developed using a minigene reporter system (AIDs, 1459 & 1487). A quinazoline (CID: 16759567) showed weak activity as an inhibitor of aberrant splicing using the minigene reporter system (AID 1498). A preliminary kinase profile of this compound showed activity in splicing related kinases belonging to the CDC-like kinase (Clk) family particularly the Clk4. The Clk family is part of the CMGC group of protein kinases which includes Dyrks and are essential kinases found in all eukaryotes. Clks have been implicated to play a role in the regulation of alternative splicing [1]; while Dyrks, in particular over expression of Dyrk1A, has been linked to diseases as Down syndrome [2] and Alzheimer disease [3]. This AID summarizes the characterization of lead Clk4 inhibitors in a panel of these related kinases: Clk1, 2 & 3, DYRK1/DYRK1A, and DYRK1B. Potency against Clk4 was also confirmed in the panel.
Protocol
Assay Protocol:
Compound IC50s were determined at Reaction Biology (http://www.reactionbiology.com) with ten concentration points starting at 10 uM in the assay using a 3-fold dilution series. The control compound staurosporine was also tested in the 10-dose IC50 mode with the same 3-fold serial dilution, again starting at 10 uM. All kinase reactions were performed using 10 uM ATP in the assay. 100% activity was taken from kinase reactions with DMSO alone (the vehicle used for compound stock solutions).
Compound IC50s were determined at Reaction Biology (http://www.reactionbiology.com) with ten concentration points starting at 10 uM in the assay using a 3-fold dilution series. The control compound staurosporine was also tested in the 10-dose IC50 mode with the same 3-fold serial dilution, again starting at 10 uM. All kinase reactions were performed using 10 uM ATP in the assay. 100% activity was taken from kinase reactions with DMSO alone (the vehicle used for compound stock solutions).
Comment
Compounds with the potency below 1uM for Clk4 are given a score of 80, the rest of them are give a score of 50.
Result Definitions
| TID | Name | Description | Histogram | Type | Unit | |
|---|---|---|---|---|---|---|
| Outcome | The BioAssay activity outcome | Outcome | ||||
| Score | The BioAssay activity ranking score |  | Integer | |||
| 1 | Phenotype | Indicates type of activity observed based on the compound activity across various assays tested. | String | |||
| 2 | Potency_Clk4 | The concentration of sample yielding half-maximal inhibition in Clk4 assay. | | Float | μM | |
| 3 | Potency_Clk1 | The concentration of sample yielding half-maximal inhibition in Clk1 assay. | | Float | μM | |
| 4 | Potency_Clk2 | The concentration of sample yielding half-maximal inhibition in Clk2 assay. | | Float | μM | |
| 5 | Potency_Clk3 | The concentration of sample yielding half-maximal inhibition in Clk3 assay. | | Float | μM | |
| 6 | Potency_Dyrk1a | The concentration of sample yielding half-maximal inhibition in Dyrk1a assay. | | Float | μM | |
| 7 | Potency_Dyrk1b | The concentration of sample yielding half-maximal inhibition in Dyrk1b assay. | | Float | μM |
Additional Information
Grant Number: 1R03MH084827-01
Data Table (Concise)
Classification
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