BC C. elegans Gene Expression ConsortiumFunded by Genome Canada and Genome British Columbia. |
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Project Objectives and Summary The objective of this project is to define the RNA expression profiles in specific tissues and cells, and developmental stages of C. elegans. Two complementary approaches are being applied: serial analysis of gene expression (SAGE), and the construction of promoter::GFP fusions for in vivo analysis of gene expression. SAGE SAGE is a sensitive and specific method for obtaining qualitative and quantitative information on expressed RNAs. SAGE will also allow us to identify non-protein-coding genes, and provide insight into alternatively spliced mRNA isoforms and their relative abundance between tissues. We are examining total mRNA populations in all developmental stages, both in whole worms and in specific cells and tissues. We have generated 17 SAGE libraries, which include all developmental stages, mutation-specific populations, and specific tissues and cells, totalling approximately 1.8 million observed tags. Tissue- and cell-specific libraries were generated from FACS-sorted cells marked by expression of specific promoter::GFP fusions. To date, we have SAGE libraries for purified embryonic muscle, gut, and a subset of neurons. Promoter::GFP fusions Monitoring in vivo expression of the fusion constucts in transgenic worms allows the determination of the developmental stage, tissue, and in some cases, the cells where a particular gene is expressed. Our goal is to build promoter::GFP fusion constructs for C. elegans genes that have human orthologues. Of the over 5000 genes that fall into this category, 2000 are being targeted by the C. elegans Gene Knockout Consortium. Fusion constructs are being created for the same set of genes, with a focus on genes expressed in muscle and nerve tissues. When coupled with SAGE and knockout data, this will provide valuable and more complete expression profiles for cells, tissues, and developmental stages. Other objectives This project includes research teams from Vancouver, Canada (listed below) and the Karolinska Institute in Sweden. |
| Members of the The Genome BC C. elegans Gene Expression Consortium: |
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PI: David Baillie Group Leader: Robert Johnsen Department of Molecular Biology and Biochemistry Simon Fraser University 8888 University Drive Burnaby, BC Canada V5A 1S6 Phone: (604) 268-6590 PI: Don Moerman GFP expression contact: Rebecca Newbury C. elegans Genomics Facility University of British Columbia 2140 - 2350 Health Sciences Mall Vancouver, BC Canada V6T 1Z3 Phone: (604) 822-3365 PIs: Marco Marra, Steven Jones Group Leader: Kim Wong C. elegans Research Group Genome Sciences Centre British Columbia Cancer Agency 600 West 10th Ave. Vancouver, BC Canada V5Z 4E6 Phone: (604) 707-5964 |
