| Cell size and cell-cycle regulation (1) |
| CG33123 | SD07726 | sp |
none
| no |
ReferenceIdentification of pathways regulating cell size and cell-cycle progression by RNAi. Bjӧrklund et al.,
2006
Many high-throughput loss-of-function analyses of the eukaryotic cell cycle have relied on the unicellular yeast species Saccharomyces cerevisiae and Schizosaccharomyces pombe. In multicellular organisms, however, additional control mechanisms regulate the cell cycle to specify the size of the organism and its constituent organs. To identify such genes, here we analysed the effect of the loss of function of 70% of Drosophila genes (including 90% of genes conserved in human) on cell-cycle progression of S2 cells using flow cytometry. To address redundancy, we also targeted genes involved in protein phosphorylation simultaneously with their homologues. We identify genes that control cell size, cytokinesis, cell death and/or apoptosis, and the G1 and G2/M phases of the cell cycle. Classification of the genes into pathways by unsupervised hierarchical clustering on the basis of these phenotypes shows that, in addition to classical regulatory mechanisms such as Myc/Max, Cyclin/Cdk and E2F, cell-cycle progression in S2 cells is controlled by vesicular and nuclear transport proteins, COP9 signalosome activity and four extracellular-signal-regulated pathways (Wnt, p38betaMAPK, FRAP/TOR and JAK/STAT). In addition, by simultaneously analysing several phenotypes, we identify a translational regulator, eIF-3p66, that specifically affects the Cyclin/Cdk pathway activity.
Screen detailsStable Id:
GR00048-A-1
Screen title:
Cell size and cell-cycle regulation (1)
Assay:
Cell size, DNA content and viability
Method:
Flow cytometry
Scope:
Kinases, phosphatases and selected genes
Screen type:
Cell-based
Species:
Drosophila melanogaster
Biosource:
Cell line
Biomodel:
S2
Library:
Custom-made, DGC1, DGC2 and PHOSPHO
Reagent type:
dsRNA
Score type:
Complex, sp
Cutoff:
Complex criteria
Notes:
Additional information about the primary sccreen (pooled library) and a secondary screen (number of binucleate cells)
|
| Lipid storage | FBgn0053123
| | | 0.03 |
none
| no |
ReferenceCOPI complex is a regulator of lipid homeostasis. Beller et al.,
2008
Lipid droplets are ubiquitous triglyceride and sterol ester storage organelles required for energy storage homeostasis and biosynthesis. Although little is known about lipid droplet formation and regulation, it is clear that members of the PAT (perilipin, adipocyte differentiation related protein, tail interacting protein of 47 kDa) protein family coat the droplet surface and mediate interactions with lipases that remobilize the stored lipids. We identified key Drosophila candidate genes for lipid droplet regulation by RNA interference (RNAi) screening with an image segmentation-based optical read-out system, and show that these regulatory functions are conserved in the mouse. Those include the vesicle-mediated Coat Protein Complex I (COPI) transport complex, which is required for limiting lipid storage. We found that COPI components regulate the PAT protein composition at the lipid droplet surface, and promote the association of adipocyte triglyceride lipase (ATGL) with the lipid droplet surface to mediate lipolysis. Two compounds known to inhibit COPI function, Exo1 and Brefeldin A, phenocopy COPI knockdowns. Furthermore, RNAi inhibition of ATGL and simultaneous drug treatment indicate that COPI and ATGL function in the same pathway. These data indicate that the COPI complex is an evolutionarily conserved regulator of lipid homeostasis, and highlight an interaction between vesicle transport systems and lipid droplets.
Screen detailsStable Id:
GR00002-A-0
Screen title:
Lipid storage
Assay:
Lipid droplet staining
Method:
High content (microscopy)
Scope:
Screen type:
Cell-based
Species:
Drosophila melanogaster
Biosource:
Cell line
Biomodel:
Kc167
Library:
, DRSC
Reagent type:
dsRNA
Score type:
B-score
Cutoff:
2.0 / -1.7
Notes:
|
| Lipid storage | FBgn0053123
| | | 0.69 |
none
| no |
ReferenceCOPI complex is a regulator of lipid homeostasis. Beller et al.,
2008
Lipid droplets are ubiquitous triglyceride and sterol ester storage organelles required for energy storage homeostasis and biosynthesis. Although little is known about lipid droplet formation and regulation, it is clear that members of the PAT (perilipin, adipocyte differentiation related protein, tail interacting protein of 47 kDa) protein family coat the droplet surface and mediate interactions with lipases that remobilize the stored lipids. We identified key Drosophila candidate genes for lipid droplet regulation by RNA interference (RNAi) screening with an image segmentation-based optical read-out system, and show that these regulatory functions are conserved in the mouse. Those include the vesicle-mediated Coat Protein Complex I (COPI) transport complex, which is required for limiting lipid storage. We found that COPI components regulate the PAT protein composition at the lipid droplet surface, and promote the association of adipocyte triglyceride lipase (ATGL) with the lipid droplet surface to mediate lipolysis. Two compounds known to inhibit COPI function, Exo1 and Brefeldin A, phenocopy COPI knockdowns. Furthermore, RNAi inhibition of ATGL and simultaneous drug treatment indicate that COPI and ATGL function in the same pathway. These data indicate that the COPI complex is an evolutionarily conserved regulator of lipid homeostasis, and highlight an interaction between vesicle transport systems and lipid droplets.
Screen detailsStable Id:
GR00002-A-0
Screen title:
Lipid storage
Assay:
Lipid droplet staining
Method:
High content (microscopy)
Scope:
Screen type:
Cell-based
Species:
Drosophila melanogaster
Biosource:
Cell line
Biomodel:
Kc167
Library:
, DRSC
Reagent type:
dsRNA
Score type:
B-score
Cutoff:
2.0 / -1.7
Notes:
|
| Heart development and function (1) | CG33123
| | | 1 |
none
| no |
ReferenceA global in vivo Drosophila RNAi screen identifies NOT3 as a conserved regulator of heart function. Neely et al.,
2010
Heart diseases are the most common causes of morbidity and death in humans. Using cardiac-specific RNAi-silencing in Drosophila, we knocked down 7061 evolutionarily conserved genes under conditions of stress. We present a first global roadmap of pathways potentially playing conserved roles in the cardiovascular system. One critical pathway identified was the CCR4-Not complex implicated in transcriptional and posttranscriptional regulatory mechanisms. Silencing of CCR4-Not components in adult Drosophila resulted in myofibrillar disarray and dilated cardiomyopathy. Heterozygous not3 knockout mice showed spontaneous impairment of cardiac contractility and increased susceptibility to heart failure. These heart defects were reversed via inhibition of HDACs, suggesting a mechanistic link to epigenetic chromatin remodeling. In humans, we show that a common NOT3 SNP correlates with altered cardiac QT intervals, a known cause of potentially lethal ventricular tachyarrhythmias. Thus, our functional genome-wide screen in Drosophila can identify candidates that directly translate into conserved mammalian genes involved in heart function.
Screen detailsStable Id:
GR00138-A-1
Screen title:
Heart development and function (1)
Assay:
Viability
Method:
Fly count
Scope:
Selected genes
Screen type:
in vivo
Species:
Drosophila melanogaster
Biosource:
Tissue
Biomodel:
TinCΔ4 12a-Gal4
Library:
VDRC, np
Reagent type:
UAS-IR construct
Score type:
Developmental lethality
Cutoff:
<= 0.6666
Notes:
|
| Notch pathway regulation (4) | CG33123
| | | sp |
Completely lethal (before pupal)
| no |
ReferenceGenome-wide analysis of Notch signalling in Drosophila by transgenic RNAi. Mummery-Widmer et al.,
2009
Genome-wide RNA interference (RNAi) screens have identified near-complete sets of genes involved in cellular processes. However, this methodology has not yet been used to study complex developmental processes in a tissue-specific manner. Here we report the use of a library of Drosophila strains expressing inducible hairpin RNAi constructs to study the Notch signalling pathway during external sensory organ development. We assigned putative loss-of-function phenotypes to 21.2% of the protein-coding Drosophila genes. Using secondary assays, we identified 6 new genes involved in asymmetric cell division and 23 novel genes regulating the Notch signalling pathway. By integrating our phenotypic results with protein interaction data, we constructed a genome-wide, functionally validated interaction network governing Notch signalling and asymmetric cell division. We used clustering algorithms to identify nuclear import pathways and the COP9 signallosome as Notch regulators. Our results show that complex developmental processes can be analysed on a genome-wide level and provide a unique resource for functional annotation of the Drosophila genome.
Screen detailsStable Id:
GR00144-A-4
Screen title:
Notch pathway regulation (4)
Assay:
External sensory organ morphology and viability
Method:
Visual inspection
Scope:
Genome-wide
Screen type:
in vivo
Species:
Drosophila melanogaster
Biosource:
Tissue
Biomodel:
pnr-GAL4
Library:
VDRC, np
Reagent type:
UAS-IR construct
Score type:
Phenotype strength
Cutoff:
np
Notes:
|
| Muscle morphogenesis and function (1) | CG33123
| CG33123 | | np |
Lethal
| no |
ReferenceSystematic genetic analysis of muscle morphogenesis and function in Drosophila. Schnorrer et al.,
2010
Systematic genetic approaches have provided deep insight into the molecular and cellular mechanisms that operate in simple unicellular organisms. For multicellular organisms, however, the pleiotropy of gene function has largely restricted such approaches to the study of early embryogenesis. With the availability of genome-wide transgenic RNA interference (RNAi) libraries in Drosophila, it is now possible to perform a systematic genetic dissection of any cell or tissue type at any stage of the lifespan. Here we apply these methods to define the genetic basis for formation and function of the Drosophila muscle. We identify a role in muscle for 2,785 genes, many of which we assign to specific functions in the organization of muscles, myofibrils or sarcomeres. Many of these genes are phylogenetically conserved, including genes implicated in mammalian sarcomere organization and human muscle diseases.
Screen detailsStable Id:
GR00134-A-1
Screen title:
Muscle morphogenesis and function (1)
Assay:
Posture, locomotion, flight and viability
Method:
Visual inspection
Scope:
Genome-wide
Screen type:
in vivo
Species:
Drosophila melanogaster
Biosource:
Tissue
Biomodel:
Mef2-GAL4
Library:
VDRC, np
Reagent type:
UAS-IR construct
Score type:
rp
Cutoff:
S19 > 0.5
Notes:
|
| Chlamydia infection | FBgn0053123
| | | np |
Less inclusions
| no |
ReferenceRNAi screen in Drosophila cells reveals the involvement of the Tom complex in Chlamydia infection. Derre et al.,
2007
Chlamydia spp. are intracellular obligate bacterial pathogens that infect a wide range of host cells. Here, we show that C. caviae enters, replicates, and performs a complete developmental cycle in Drosophila SL2 cells. Using this model system, we have performed a genome-wide RNA interference screen and identified 54 factors that, when depleted, inhibit C. caviae infection. By testing the effect of each candidate''s knock down on L. monocytogenes infection, we have identified 31 candidates presumably specific of C. caviae infection. We found factors expected to have an effect on Chlamydia infection, such as heparansulfate glycosaminoglycans and actin and microtubule remodeling factors. We also identified factors that were not previously described as involved in Chlamydia infection. For instance, we identified members of the Tim-Tom complex, a multiprotein complex involved in the recognition and import of nuclear-encoded proteins to the mitochondria, as required for C. caviae infection of Drosophila cells. Finally, we confirmed that depletion of either Tom40 or Tom22 also reduced C. caviae infection in mammalian cells. However, C. trachomatis infection was not affected, suggesting that the mechanism involved is C. caviae specific.
Screen detailsStable Id:
GR00013-A-0
Screen title:
Chlamydia infection
Assay:
Chlamydia infection
Method:
High content (microscopy)
Scope:
Screen type:
Cell-based
Species:
Drosophila melanogaster
Biosource:
Cell line
Biomodel:
SL2
Library:
, DRSC
Reagent type:
dsRNA
Score type:
Visual inspection
Cutoff:
np
Notes:
|
| Self-renewal and differentiation in neural stem cells | FBgn0053123
| CG33123 | | sp |
GFP aggregates number, GFP aggregates size, neuroblast smaller cell size, neuroblast less large cells, neuroblast cell shape, ganglion mother cell shorter lineages, ganglion mother cell shape, neuroblast loss, neuroblast loss with smaller neuroblast cell
| no |
ReferenceGenome-wide analysis of self-renewal in Drosophila neural stem cells by transgenic RNAi. Neumueller et al.,
2011
The balance between stem cell self-renewal and differentiation is precisely controlled to ensure tissue homeostasis and prevent tumorigenesis. Here we use genome-wide transgenic RNAi to identify 620 genes potentially involved in controlling this balance in Drosophila neuroblasts. We quantify all phenotypes and derive measurements for proliferation, lineage, cell size, and cell shape. We identify a set of transcriptional regulators essential for self-renewal and use hierarchical clustering and integration with interaction data to create functional networks for the control of neuroblast self-renewal and differentiation. Our data identify key roles for the chromatin remodeling Brm complex, the spliceosome, and the TRiC/CCT-complex and show that the alternatively spliced transcription factor Lola and the transcriptional elongation factors Ssrp and Barc control self-renewal in neuroblast lineages. As our data are strongly enriched for genes highly expressed in murine neural stem cells, they are likely to provide valuable insights into mammalian stem cell biology as well.
Screen detailsStable Id:
GR00183-A
Screen title:
Self-renewal and differentiation in neural stem cells
Assay:
Number and size of neuroblasts, ganglion mother cells, intracellular GFP aggregates and viability
Method:
Fluorescence
Scope:
Genome-wide
Screen type:
in vivo
Species:
Drosophila melanogaster
Biosource:
Tissue
Biomodel:
insc-GAL4
Library:
VDRC, np
Reagent type:
UAS-IR construct
Score type:
Phenotype strength
Cutoff:
> 1
Notes:
Additional information about a secondary screen (KK library)
|
| Viability after serine protease inactivation (2) | CG3229
| SP107 | CG3229 | np |
Viable
| yes |
ReferenceDrosophila immunity: a large-scale in vivo RNAi screen identifies five serine proteases required for Toll activation. Kambris et al.,
2006
Unlike mammalian Toll-like Receptors, the Drosophila Toll receptor does not interact directly with microbial determinants but is rather activated upon binding a cleaved form of the cytokine-like molecule Spatzle (Spz). During the immune response, Spz is thought to be processed by secreted serine proteases (SPs) present in the hemolymph that are activated by the recognition of gram-positive bacteria or fungi . In the present study, we have used an in vivo RNAi strategy to inactivate 75 distinct Drosophila SP genes. We then screened this collection for SPs regulating the activation of the Toll pathway by gram-positive bacteria. Here, we report the identification of five novel SPs that function in an extracellular pathway linking the recognition proteins GNBP1 and PGRP-SA to Spz. Interestingly, four of these genes are also required for Toll activation by fungi, while one is specifically associated with signaling in response to gram-positive bacterial infections. These results demonstrate the existence of a common cascade of SPs upstream of Spz, integrating signals sent by various secreted recognition molecules via more specialized SPs.
Screen detailsStable Id:
GR00146-A-2
Screen title:
Viability after serine protease inactivation (2)
Assay:
Viability
Method:
Fly count
Scope:
Selected genes
Screen type:
in vivo
Species:
Drosophila melanogaster
Biosource:
Tissue
Biomodel:
c564-GAL4
Library:
Custom-made, Custom-made
Reagent type:
UAS-IR construct
Score type:
Number
Cutoff:
Representative of >= 3 tests
Notes:
|
| Toll activation by gram-positive bacteria (2) | CG3229
| SP107 | CG3229 | np |
none
| no |
ReferenceDrosophila immunity: a large-scale in vivo RNAi screen identifies five serine proteases required for Toll activation. Kambris et al.,
2006
Unlike mammalian Toll-like Receptors, the Drosophila Toll receptor does not interact directly with microbial determinants but is rather activated upon binding a cleaved form of the cytokine-like molecule Spatzle (Spz). During the immune response, Spz is thought to be processed by secreted serine proteases (SPs) present in the hemolymph that are activated by the recognition of gram-positive bacteria or fungi . In the present study, we have used an in vivo RNAi strategy to inactivate 75 distinct Drosophila SP genes. We then screened this collection for SPs regulating the activation of the Toll pathway by gram-positive bacteria. Here, we report the identification of five novel SPs that function in an extracellular pathway linking the recognition proteins GNBP1 and PGRP-SA to Spz. Interestingly, four of these genes are also required for Toll activation by fungi, while one is specifically associated with signaling in response to gram-positive bacterial infections. These results demonstrate the existence of a common cascade of SPs upstream of Spz, integrating signals sent by various secreted recognition molecules via more specialized SPs.
Screen detailsStable Id:
GR00145-A-2
Screen title:
Toll activation by gram-positive bacteria (2)
Assay:
Drosomycin mRNA expression
Method:
Real-time qPCR
Scope:
Selected genes
Screen type:
in vivo
Species:
Drosophila melanogaster
Biosource:
Organism
Biomodel:
da-GAL4 AND c564-GAL4
Library:
Custom-made, Custom-made
Reagent type:
UAS-IR construct
Score type:
Percentage
Cutoff:
> 50% decrease
Notes:
|
| Viability after serine protease inactivation (1) | CG3229
| SP107 | CG3229 | np |
Lethal only with R2 insertion
| yes |
ReferenceDrosophila immunity: a large-scale in vivo RNAi screen identifies five serine proteases required for Toll activation. Kambris et al.,
2006
Unlike mammalian Toll-like Receptors, the Drosophila Toll receptor does not interact directly with microbial determinants but is rather activated upon binding a cleaved form of the cytokine-like molecule Spatzle (Spz). During the immune response, Spz is thought to be processed by secreted serine proteases (SPs) present in the hemolymph that are activated by the recognition of gram-positive bacteria or fungi . In the present study, we have used an in vivo RNAi strategy to inactivate 75 distinct Drosophila SP genes. We then screened this collection for SPs regulating the activation of the Toll pathway by gram-positive bacteria. Here, we report the identification of five novel SPs that function in an extracellular pathway linking the recognition proteins GNBP1 and PGRP-SA to Spz. Interestingly, four of these genes are also required for Toll activation by fungi, while one is specifically associated with signaling in response to gram-positive bacterial infections. These results demonstrate the existence of a common cascade of SPs upstream of Spz, integrating signals sent by various secreted recognition molecules via more specialized SPs.
Screen detailsStable Id:
GR00146-A-1
Screen title:
Viability after serine protease inactivation (1)
Assay:
Viability
Method:
Fly count
Scope:
Serine proteases
Screen type:
in vivo
Species:
Drosophila melanogaster
Biosource:
Organism
Biomodel:
da-GAL4
Library:
Custom-made, Custom-made
Reagent type:
UAS-IR construct
Score type:
Number
Cutoff:
Representative of >= 3 tests
Notes:
|
| Heat nociception (1) | CG3229
| CG33123 | | -1.17 |
none
| yes |
ReferenceA genome-wide Drosophila screen for heat nociception identifies α2δ3 as an evolutionarily conserved pain gene. Neely et al.,
2010
Worldwide, acute, and chronic pain affects 20% of the adult population and represents an enormous financial and emotional burden. Using genome-wide neuronal-specific RNAi knockdown in Drosophila, we report a global screen for an innate behavior and identify hundreds of genes implicated in heat nociception, including the α2δ family calcium channel subunit straightjacket (stj). Mice mutant for the stj ortholog CACNA2D3 (α2δ3) also exhibit impaired behavioral heat pain sensitivity. In addition, in humans, α2δ3 SNP variants associate with reduced sensitivity to acute noxious heat and chronic back pain. Functional imaging in α2δ3 mutant mice revealed impaired transmission of thermal pain-evoked signals from the thalamus to higher-order pain centers. Intriguingly, in α2δ3 mutant mice, thermal pain and tactile stimulation triggered strong cross-activation, or synesthesia, of brain regions involved in vision, olfaction, and hearing.
Screen detailsStable Id:
GR00135-A-1
Screen title:
Heat nociception (1)
Assay:
Noxious heat avoidance and viability
Method:
Fly count
Scope:
Genome-wide
Screen type:
in vivo
Species:
Drosophila melanogaster
Biosource:
Organism
Biomodel:
elav-GAL4
Library:
VDRC, np
Reagent type:
UAS-IR construct
Score type:
Z-score
Cutoff:
> 1.65
Notes:
Additional information about secondary screens (geotactic, phototaxis, and temperature sensitivity)
|
| Cell growth and viability (2) |
| | | -0.5 |
none
| no |
ReferenceGenome-wide RNAi analysis of growth and viability in Drosophila cells. Boutros et al.,
2004
A crucial aim upon completion of whole genome sequences is the functional analysis of all predicted genes. We have applied a high-throughput RNA-interference (RNAi) screen of 19,470 double-stranded (ds) RNAs in cultured cells to characterize the function of nearly all (91%) predicted Drosophila genes in cell growth and viability. We found 438 dsRNAs that identified essential genes, among which 80% lacked mutant alleles. A quantitative assay of cell number was applied to identify genes of known and uncharacterized functions. In particular, we demonstrate a role for the homolog of a mammalian acute myeloid leukemia gene (AML1) in cell survival. Such a systematic screen for cell phenotypes, such as cell viability, can thus be effective in characterizing functionally related genes on a genome-wide scale.
Screen detailsStable Id:
GR00031-A-2
Screen title:
Cell growth and viability (2)
Assay:
Cell number and viability
Method:
Luminescence
Scope:
Genome-wide
Screen type:
Cell-based
Species:
Drosophila melanogaster
Biosource:
Cell line
Biomodel:
S2R+
Library:
Custom-made, HFA
Reagent type:
dsRNA
Score type:
Z-score
Cutoff:
>= 3.0
Notes:
|
| Muscle morphogenesis and function (1) | CG33123
| CG33123 | | np |
none
| no |
ReferenceSystematic genetic analysis of muscle morphogenesis and function in Drosophila. Schnorrer et al.,
2010
Systematic genetic approaches have provided deep insight into the molecular and cellular mechanisms that operate in simple unicellular organisms. For multicellular organisms, however, the pleiotropy of gene function has largely restricted such approaches to the study of early embryogenesis. With the availability of genome-wide transgenic RNA interference (RNAi) libraries in Drosophila, it is now possible to perform a systematic genetic dissection of any cell or tissue type at any stage of the lifespan. Here we apply these methods to define the genetic basis for formation and function of the Drosophila muscle. We identify a role in muscle for 2,785 genes, many of which we assign to specific functions in the organization of muscles, myofibrils or sarcomeres. Many of these genes are phylogenetically conserved, including genes implicated in mammalian sarcomere organization and human muscle diseases.
Screen detailsStable Id:
GR00134-A-1
Screen title:
Muscle morphogenesis and function (1)
Assay:
Posture, locomotion, flight and viability
Method:
Visual inspection
Scope:
Genome-wide
Screen type:
in vivo
Species:
Drosophila melanogaster
Biosource:
Tissue
Biomodel:
Mef2-GAL4
Library:
VDRC, np
Reagent type:
UAS-IR construct
Score type:
rp
Cutoff:
S19 > 0.5
Notes:
|
| Heart development and function (1) | CG33123
| | | 0.83 |
none
| no |
ReferenceA global in vivo Drosophila RNAi screen identifies NOT3 as a conserved regulator of heart function. Neely et al.,
2010
Heart diseases are the most common causes of morbidity and death in humans. Using cardiac-specific RNAi-silencing in Drosophila, we knocked down 7061 evolutionarily conserved genes under conditions of stress. We present a first global roadmap of pathways potentially playing conserved roles in the cardiovascular system. One critical pathway identified was the CCR4-Not complex implicated in transcriptional and posttranscriptional regulatory mechanisms. Silencing of CCR4-Not components in adult Drosophila resulted in myofibrillar disarray and dilated cardiomyopathy. Heterozygous not3 knockout mice showed spontaneous impairment of cardiac contractility and increased susceptibility to heart failure. These heart defects were reversed via inhibition of HDACs, suggesting a mechanistic link to epigenetic chromatin remodeling. In humans, we show that a common NOT3 SNP correlates with altered cardiac QT intervals, a known cause of potentially lethal ventricular tachyarrhythmias. Thus, our functional genome-wide screen in Drosophila can identify candidates that directly translate into conserved mammalian genes involved in heart function.
Screen detailsStable Id:
GR00138-A-1
Screen title:
Heart development and function (1)
Assay:
Viability
Method:
Fly count
Scope:
Selected genes
Screen type:
in vivo
Species:
Drosophila melanogaster
Biosource:
Tissue
Biomodel:
TinCΔ4 12a-Gal4
Library:
VDRC, np
Reagent type:
UAS-IR construct
Score type:
Developmental lethality
Cutoff:
<= 0.6666
Notes:
|
| Regulated secretory pathway biogenesis (1) | CG33123
| | np | 0.4 |
none
| no |
ReferenceRNAi screen identifies a role for adaptor protein AP-3 in sorting to the regulated secretory pathway. Asensio et al.,
2010
The regulated release of proteins depends on their inclusion within large dense-core vesicles (LDCVs) capable of regulated exocytosis. LDCVs form at the trans-Golgi network (TGN), but the mechanism for protein sorting to this regulated secretory pathway (RSP) and the cytosolic machinery involved in this process have remained poorly understood. Using an RNA interference screen in Drosophila melanogaster S2 cells, we now identify a small number of genes, including several subunits of the heterotetrameric adaptor protein AP-3, which are required for sorting to the RSP. In mammalian neuroendocrine cells, loss of AP-3 dysregulates exocytosis due to a primary defect in LDCV formation. Previous work implicated AP-3 in the endocytic pathway, but we find that AP-3 promotes sorting to the RSP within the biosynthetic pathway at the level of the TGN. Although vesicles with a dense core still form in the absence of AP-3, they contain substantially less synaptotagmin 1, indicating that AP-3 concentrates the proteins required for regulated exocytosis.
Screen detailsStable Id:
GR00137-A-1
Screen title:
Regulated secretory pathway biogenesis (1)
Assay:
dVMAT protein cell surface expression
Method:
Flow cytometry
Scope:
Selected genes
Screen type:
Cell-based
Species:
Drosophila melanogaster
Biosource:
Cell line
Biomodel:
S2
Library:
University of California San Francisco, DmRNAi library version 1
Reagent type:
dsRNA
Score type:
Z-score
Cutoff:
>= 3.0
Notes:
|
| Cell growth and viability (1) |
| | | -0.7 |
none
| yes |
ReferenceGenome-wide RNAi analysis of growth and viability in Drosophila cells. Boutros et al.,
2004
A crucial aim upon completion of whole genome sequences is the functional analysis of all predicted genes. We have applied a high-throughput RNA-interference (RNAi) screen of 19,470 double-stranded (ds) RNAs in cultured cells to characterize the function of nearly all (91%) predicted Drosophila genes in cell growth and viability. We found 438 dsRNAs that identified essential genes, among which 80% lacked mutant alleles. A quantitative assay of cell number was applied to identify genes of known and uncharacterized functions. In particular, we demonstrate a role for the homolog of a mammalian acute myeloid leukemia gene (AML1) in cell survival. Such a systematic screen for cell phenotypes, such as cell viability, can thus be effective in characterizing functionally related genes on a genome-wide scale.
Screen detailsStable Id:
GR00031-A-1
Screen title:
Cell growth and viability (1)
Assay:
Cell number and viability
Method:
Luminescence
Scope:
Genome-wide
Screen type:
Cell-based
Species:
Drosophila melanogaster
Biosource:
Cell line
Biomodel:
Kc167
Library:
Custom-made, HFA
Reagent type:
dsRNA
Score type:
Z-score
Cutoff:
>= 3.0
Notes:
|
| Toll activation by gram-positive bacteria (1) | CG3229
| SP107 | CG3229 | np |
Moderately increased Enterococcus faecalis infection
| yes |
ReferenceDrosophila immunity: a large-scale in vivo RNAi screen identifies five serine proteases required for Toll activation. Kambris et al.,
2006
Unlike mammalian Toll-like Receptors, the Drosophila Toll receptor does not interact directly with microbial determinants but is rather activated upon binding a cleaved form of the cytokine-like molecule Spatzle (Spz). During the immune response, Spz is thought to be processed by secreted serine proteases (SPs) present in the hemolymph that are activated by the recognition of gram-positive bacteria or fungi . In the present study, we have used an in vivo RNAi strategy to inactivate 75 distinct Drosophila SP genes. We then screened this collection for SPs regulating the activation of the Toll pathway by gram-positive bacteria. Here, we report the identification of five novel SPs that function in an extracellular pathway linking the recognition proteins GNBP1 and PGRP-SA to Spz. Interestingly, four of these genes are also required for Toll activation by fungi, while one is specifically associated with signaling in response to gram-positive bacterial infections. These results demonstrate the existence of a common cascade of SPs upstream of Spz, integrating signals sent by various secreted recognition molecules via more specialized SPs.
Screen detailsStable Id:
GR00145-A-1
Screen title:
Toll activation by gram-positive bacteria (1)
Assay:
Viability
Method:
Fly count
Scope:
Selected genes
Screen type:
in vivo
Species:
Drosophila melanogaster
Biosource:
Organism
Biomodel:
da-GAL4 AND c564-GAL4
Library:
Custom-made, Custom-made
Reagent type:
UAS-IR construct
Score type:
Number
Cutoff:
Representative of >= 3 tests
Notes:
|
| Notch pathway regulation (4) | CG33123
| | | 0 |
none
| no |
ReferenceGenome-wide analysis of Notch signalling in Drosophila by transgenic RNAi. Mummery-Widmer et al.,
2009
Genome-wide RNA interference (RNAi) screens have identified near-complete sets of genes involved in cellular processes. However, this methodology has not yet been used to study complex developmental processes in a tissue-specific manner. Here we report the use of a library of Drosophila strains expressing inducible hairpin RNAi constructs to study the Notch signalling pathway during external sensory organ development. We assigned putative loss-of-function phenotypes to 21.2% of the protein-coding Drosophila genes. Using secondary assays, we identified 6 new genes involved in asymmetric cell division and 23 novel genes regulating the Notch signalling pathway. By integrating our phenotypic results with protein interaction data, we constructed a genome-wide, functionally validated interaction network governing Notch signalling and asymmetric cell division. We used clustering algorithms to identify nuclear import pathways and the COP9 signallosome as Notch regulators. Our results show that complex developmental processes can be analysed on a genome-wide level and provide a unique resource for functional annotation of the Drosophila genome.
Screen detailsStable Id:
GR00144-A-4
Screen title:
Notch pathway regulation (4)
Assay:
External sensory organ morphology and viability
Method:
Visual inspection
Scope:
Genome-wide
Screen type:
in vivo
Species:
Drosophila melanogaster
Biosource:
Tissue
Biomodel:
pnr-GAL4
Library:
VDRC, np
Reagent type:
UAS-IR construct
Score type:
Phenotype strength
Cutoff:
np
Notes:
|
| S2 cell spreading | CG33123
| CG33123 | np | np |
none
| no |
ReferenceA whole genome RNAi screen of Drosophila S2 cell spreading performed using automated computational image analysis. D'Ambrosio and Vale,
2010
Recent technological advances in microscopy have enabled cell-based whole genome screens, but the analysis of the vast amount of image data generated by such screens usually proves to be rate limiting. In this study, we performed a whole genome RNA interference (RNAi) screen to uncover genes that affect spreading of Drosophila melanogaster S2 cells using several computational methods for analyzing the image data in an automated manner. Expected genes in the Scar-Arp2/3 actin nucleation pathway were identified as well as casein kinase I, which had a similar morphological RNAi signature. A distinct nonspreading morphological phenotype was identified for genes involved in membrane secretion or synthesis. In this group, we identified a new secretory peptide and investigated the functions of two poorly characterized endoplasmic reticulum proteins that have roles in secretion. Thus, this genome-wide screen succeeded in identifying known and unexpected proteins that are important for cell spreading, and the computational tools developed in this study should prove useful for other types of automated whole genome screens.
Screen detailsStable Id:
GR00194-A
Screen title:
S2 cell spreading
Assay:
alpha-tubulin and actin protein expression
Method:
Fluorescence
Scope:
Genome-wide
Screen type:
Cell-based
Species:
Drosophila melanogaster
Biosource:
Cell line
Biomodel:
S2U
Library:
Thermo Fisher Scientific, V2 RNAi library
Reagent type:
dsRNA
Score type:
Complex, sp
Cutoff:
Complex criteria
Notes:
|
