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BioAssay: AID 686938

Genes involved in store-operated calcium entry

Store-operated Ca2+ entry is mediated by Ca2+ release-activated Ca2+ (CRAC) channels following Ca2+ release from intracellular stores. We performed a genomewide RNA interference (RNAi) screen in Drosophila cells to identify proteins that inhibit store-operated Ca2+ influx. A secondary patch-clamp screen identified CRACM1 and CRACM2 (CRAC modulators 1 and 2) as modulators of Drosophila CRAC more ..
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 Tested RNAi Reagents
 Tested RNAi Reagents
All(22913)
 
 
Active(75)
 
 
Inactive(22838)
 
 
 Screened Genes
 Screened Genes
AID: 686938
Data Source: Drosophila RNAi Screening Center (DRSC) (DRSC-P54)
BioAssay Type: Primary, Primary Screening, Single Concentration Activity Observed
Depositor Category: Assay Vendor
Deposit Date: 2013-04-13

Data Table ( Complete ):           View Active Data    View All Data
Description:
Title: CRACM1 Is a Plasma Membrane Protein Essential for Store-Operated Ca2+ Entry

Abstract:
Store-operated Ca2+ entry is mediated by Ca2+ release-activated Ca2+ (CRAC) channels following Ca2+ release from intracellular stores. We performed a genomewide RNA interference (RNAi) screen in Drosophila cells to identify proteins that inhibit store-operated Ca2+ influx. A secondary patch-clamp screen identified CRACM1 and CRACM2 (CRAC modulators 1 and 2) as modulators of Drosophila CRAC currents. We characterized the human ortholog of CRACM1, a plasma membrane-resident protein encoded by gene FLJ14466. Although overexpression of CRACM1 did not affect CRAC currents, RNAi-mediated knockdown disrupted its activation. CRACM1 could be the CRAC channel itself, a subunit of it, or a component of the CRAC signaling machinery.
Protocol
To identify genes encoding the CRAC channel or other proteins involved in its regulation, we performed a high-throughput, genome-wide RNA interference (RNAi) screen in Drosophila S2R+ cells. The effect of knockdown of each of the ~23,000 genes was tested by fluorescence measurements of intracellular Ca2+ concentration in 384-well microplates with an automated fluorometric imaging plate reader (FLIPR, Molecular Devices). Changes in [Ca2+]i were measured in response to the commonly used SERCA (sarco-endoplasmic reticulum calcium ATPase) inhibitor thapsigargin which causes depletion of Ca2+ from intracellular stores. All 63 plates contained wells in which double-stranded RNA (dsRNA) against Rho1 served as negative control and dsRNA against stim1 as positive control.
Comment
The activity scores are given using the standard DRSC scoring system for hits:
1 = weak hit
2 = medium hit
3 = strong hit
In many assays, only 2 is used.
Result Definitions
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TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1basal replicate 1Float
2basal replicate 2Float
3basal replicate 3Float
4basal replicate 4Float
5basal replicate 5Float
6basal replicate 6Float
7basal replicate 7Float
8basal replicate 8Float
9CCE replicate 1Float
10CCE replicate 2Float
11CCE replicate 3Float
12CCE replicate 4Float
13CCE replicate 5Float
14CCE replicate 6Float
15CCE replicate 7Float
16CCE replicate 8Float
17Fmax replicate 1Float
18Fmax replicate 2Float
19Fmax replicate 3Float
20Fmax replicate 4Float
21Fmax replicate 5Float
22Fmax replicate 6Float
23Fmax replicate 7Float
24Fmax replicate 8Float
25CCE/basalFloat
26Predicted Target GeneString

RNAi Target.
Additional Information
Substance Type: Nucleotide

Data Table (Concise)
Data Table ( Complete ):     View Active Data    View All Data
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