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
Genetic Approaches | Gene Screens | Chemical Screens
Genetic Approaches
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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) |