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Discovery - Activities

Objectives

  • to use genetic and expression strategies in genetically tractable organisms to identify migration related genes
  • to use expression cloning strategies to identify proteins involved in cell migration
  • to screen chemical libraries for reagents that perturb migration, adhesion or cytoskeletal organization

Discovery Protocols

Genetic Approaches | Gene Screens | Chemical Screens

Genetic Approaches

Organism Approach Status / Publication Contact (Collaborators)
C. elegans Determine cell autonomy of 99 DTC migration genes Identified 28 genes with cell autonomous functions in distal tip cell migration and gonadogenesis
Data available		 Schwarzbauer
C. elegans RNAi screen for DTC migration defects Analysis of 16,758 RNA-interference depletion experiments identified 99 genes required for distal tip cell migration
Data available		 Schwarzbauer
D. melanogaster characterization of the proteins identified in the high throughput screen Completed - Contact PI Vale
D. melanogaster Collection of published cell migration genes in Drosophila (multiple cell types) Rac, Shg and Stat92e are implicated in both primordial germ cell and in border cell migration
Data available		 Montell
D. melanogaster identification of downstream targets of migration-related transcription factors Identified 149 genes expressed at a lower level in border cells isolated from slbo mutants
Data available		 Montell
D. melanogaster identification of genes encoding cytoskeleton-associated proteins enriched in migratory cells 45 genes enriched in migratory cells
Data available		 Montell
D. melanogaster identification of genes that alter S2 morphology using large scale RNAi screening and high resolution/high-throughput microscopy Data available - D'Ambrosio and Vale, 2010 (CMC Update Article) Vale
D. melanogaster identification of genes upregulated in migrating cells 2736 examined, genes predicted to control actin dynamics and endocytic and secretory pathways are overrepresented in the migratory cell transcriptome
Data available		 Montell
D. melanogaster identification of migratory cell enriched genes encoding proteins associated with vesicle traffic 46 genes enriched in migratory cells, encoding vesicle associated proteins
Data available		 Montell
D. melanogaster matching border cell genes with migration defective mutants Completed - Contact PI Montell
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Gene Screens

Phenotype Screen Target Cell Library (cDNA / siRNA) Status / Publication Contact (Collaborators)
Invasive Cell translocation in response to metastasis signal PyMT and MTLn3 Genes identified characteristic of the "invasion signature"
Completed - Data available		 Condeelis
Migration Phagokinetic track assay (PKT) MCF-7 BC-1000 cDNA 4 novel genes identified in BC1000 screen that alter migration of the MCF7 cells
Completed - Data available - Naffar-Abu-Amara et al., 2008 (CMC Update Article)		 Geiger (Naffar-Abu-Amara, Brugge)
Migration Phagokinetic track assay (PKT) MCF-7 MDA-MB-231 cDNA Paper submitted for publication, data available upon request
Completed - Data available		 Geiger (Naffar-Abu-Amara, Brugge)
Migration Phagokinetic track assay (PKT-B) MCF-10A siRNA-kinases, phosphatases and custom set Effects of siRNA-induced gene knockdown include decrease in focal adhesion number and change in distribution throughout the cell
Completed - Data available		 Geiger
Migration Transwell assay for induction of migration MCF-10A MCF-7 cDNA 6 genes identified that increase migratory behavior
Completed - Data available		 Brugge
Migration Transwell assay for induction of migration MCF-10A - (expressing activated ErbB2) BC-1000 cDNA 39 out of 41 genes increase migratory behavior in MCF-10A expressing activated ErbB2
Completed - Data available		 Brugge (LaBaer)
Migration Transwell assay for induction of migration MCF-10A - (expressing activated ErbB2) MCF-7 cDNA 3 genes identified that increase migratory behavior in MCF-10A expressing activated ErbB2
Completed - Data available		 Brugge
Migration Transwell assay for induction of migration MCF-10A - (expressing CSF-1R and stimulated with CSF) MCF-7 cDNA 4 genes identified that increase migratory behavior in MCF-10A expressing CSF-1R and stimulated with CSF
Completed - Data available		 Brugge
Migration Transwell assay for induction of migration MCF-10A - (expressing CSF-1R) MCF-7 cDNA 4 genes identified that increase migratory behavior in MCF-10A expressing CSF-1R
Completed - Data available		 Brugge
Migration Transwell assay for induction of migration in the absence of a functional PI3K pathway MCF-10A - (stimulated with EGF + PI3K inhibition with LY294002) MDA-MB-231 cDNA Completed - no hits Brugge
Migration Wound healing assay MCF-10A siRNA-kinases, phosphatases and custom set 66 genes identified that positively or negatively regulate cell migration
Completed - Data available - Simpson et al., 2008 (CMC Update Article)		 Brugge
Structure Cytoskeletal localization REF52 YFP-tagged rat cDNA The localization to migratory structures has been identified for 202 clones
Completed - Data available		 Geiger (Lavelin)
Structure Cytoskeletal localization REF52 YFP-tagged arabidopsis cDNA Identified ERD10 associated with actin stress fibers, RC-GAP72 plays a role in regulating cell morphology and cytoskeletal organization.
Completed - Lavelin and Geiger, 2005; Abu-Abied et al., 2006; Paran et al., 2006 (CMC Update Article)		 Geiger (Sadot, Kam)
Structure Localization REF52 GFP-tagged HT1080 cDNA clones identified that localize to migration related structures
Completed - Data available - Manabe et al., 2002 (CMC Update Article)		 Horwitz
Structure Morphological perturbations HeLa with paxillin-YFP reporter siRNA-kinases Effect gene knockdown of 576 kinases on the morphology of focal adhesions is shown
Completed - Data available		 Geiger (Winograd-Katz, Brugge)
Structure Morphological perturbations HeLa with paxillin-YFP reporter siRNA-phosphatases Effect of gene knockdown of 192 phosphatases on the morphology of focal adhesions is shown
Completed - Data available		 Geiger (Winograd-Katz, Brugge)
Structure Morphological perturbations HeLa with paxillin-YFP reporter siRNA-Migration and Adhesion related genes Effect of knockdown of 312 Migration-related genes on the morphology of focal adhesion is shown
Completed - Data available		 Geiger (Winograd-Katz, Brugge)

Methods for validation of siRNA targeting sequences - Gary Borisy.

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Chemical Screens

Phenotype Screen Target Cell Library Status / Publication Contact (Collaborators)
Structure Morphological perturbations REF52 with YFP paxillin Tel Aviv HTS Natural Products 17 compounds identified that perturb migration-related structures
Completed - Data available		 Geiger (Paran)
Structure Morphological perturbations REF52 with YFP paxillin Chem Bridge & Chemical Diversity 102 compounds identified that perturb migration-related structures
Completed - Data available		 Geiger (Paran)
Structure Phagokinetic track assay (PKT) MCF-7 / MDA-231-MB Tel Aviv HTS Natural Products Completed - Contact PI Geiger (Naffar-Abu-Amara)
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