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BioAssay: AID 2598

qHTS Assay for General miRNAs Modulators and/or Inhibitors/Activators of miR-21: Summary

MicroRNAs (miRNAs) are endogenously encoded small (~20-25 nucleotides), nonprotein-coding RNAs that are involved in post-transcriptional repression of target messenger RNAs (mRNAs) (Le and Hannon, 2004). Estimated to be involved in the regulation of 30% of all protein-coding mRNAs (Filipowicz et al., 2008), miRNAs play a significant role in many biological processes including cellular more ..
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 Related BioAssays
 Related BioAssays
AID: 2598
Data Source: NCGC (MIRNA396)
BioAssay Type: Summary, Candidate Probes/Leads with Supporting Evidence
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2010-03-18
Modify Date: 2010-03-20
Target
Depositor Specified Assays
AIDNameTypeComment
2288qHTS Assay for Modulators of miRNAs and/orActivators of miR-21confirmatoryPrimary Assay for General miRNAs Modulators and/or Activators of miR-21
2289qHTS Assay for Modulators of miRNAs and/or Inhibitors of miR-21confirmatoryPrimary Assay for General miRNAs Modulators and/or Inhibitors of miR-21
2507Secondary Assay for General miRNAs Modulators and/or Inhibitors/Activators of miR-21: miR-30 Firefly Luciferase AssayconfirmatorySecondary Assay for General miRNAs Modulators and/or Inhibitors/Activators of miR-21: miR-30 assay
2508Confirmatory Assay for General miRNAs Modulators and/or Inhibitors/Activators of miR-21confirmatoryConfirmatory Assay for General miRNAs Modulators and/or Inhibitors/Activators of miR-21
588342qHTS profiling assay for firefly luciferase inhibitor/activator using purifed enzyme and Km concentrations of substrates (counterscreen for miR-21 project)confirmatory
493175miR-21 counterscreen using purified firefly luciferaseconfirmatory
493179Secondary Assay for General miRNAs Modulators and/or Inhibitors/Activators of miR-21: miR-30 Firefly Luciferase Assay for second round cherrypicksconfirmatory
493174Confirmatory Assay for General miRNAs Modulators and/or Inhibitors/Activators of miR-21: second round cherrypicksconfirmatory
Description:
qHTS Assay for General miRNAs Modulators and/or Inhibitors/Activators of miR-21

MLSCN Grant: 1 R21 NS059478-01

Assay Provider: Qihong Huang, The Wistar Institute

NCGC Assay Overview:

MicroRNAs (miRNAs) are endogenously encoded small (~20-25 nucleotides), nonprotein-coding RNAs that are involved in post-transcriptional repression of target messenger RNAs (mRNAs) (Le and Hannon, 2004). Estimated to be involved in the regulation of 30% of all protein-coding mRNAs (Filipowicz et al., 2008), miRNAs play a significant role in many biological processes including cellular differentiation and tissue development (Ghildiyal and Zamore, 2009). Not surprisingly, miRNAs have been found to be oncogenic or tumor suppressive, with altered miRNA expression levels associated with human cancers (Esquela-Kerscher and Slack, 2006). The miRNA miR-21 has been found to be expressed in most human tissues (Sonkoly et al., 2007), and its overexpression has been associated with a variety of human tumors (Tong and Nemunanitis, 2008). miR-21 has been shown to be anti-apoptotic: knock-down of miR-21 in a glioblastoma cell line was found to activate caspases and lead to apoptosis (Chan et al., 2005). Identification of small molecules that modulate miR-21 specifically, or miRNAs generally, would prove useful as tools to better understand miRNA biogenesis and mRNA regulation by miRNAs. Of special interest are compounds that specifically inhibit miR-21, and a cell-based firefly luciferase reporter gene assay optimized for 1536-well format quantitative HTS (qHTS) has been developed in an effort to identify such compounds. The primary qHTS and confirmatory data are available on PubChem (AIDs: 2288, 2289, and 2508). A secondary assay to determine if compounds active in the miR-21 FLuc assay were potentially specific for miR-21 and were not general modulators of the miRNA pathway was another cell-based FLuc reporter gene assay, this time targeting miR-30. Data for the miR-30 FLuc assay is also available (PubChem AID 2507). Compounds found active in both the miR-21 FLuc and miR-30 FLuc assays are not considered miR-21 specific compounds, and may be general miRNA modulators.
Protocol
Please refer to other AIDs (2288, 2289, 2507, 2508) for detailed assay protocols.
Comment
This summary is written for the purposes of summarizing the probe activities from the project. We will update this summary assay once we have data available.
Additional Information
Grant Number: 1 R21 NS059478-01

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