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Comments on Ho, Margaret et al. (2013) International Worm Meeting "Mapping transcriptional regulatory networks in the nematode Caenorhabditis elegans." (0)
Overview
Ho, Margaret, & Sternberg, Paul (2013). Mapping transcriptional regulatory networks in the nematode Caenorhabditis elegans presented in International Worm Meeting. Unpublished information; cite only with author permission.
The nematode Caenorhabditis elegans has a well-annotated genome, well-studied development and many genetic tools available. C. elegans provides an excellent case to study transcriptional regulation within a multicellular organism over its entire lifecycle, especially as it is easy to collect synchronized populations of worms in distinct developmental stages. In eukaryotes, transcriptional regulation of gene expression is controlled by cis-regulatory modules (CRMs) located within non-coding regions of the genome. These CRMs include enhancers, repressors and insulators. Conventional methods in this nematode to identify CRMs frequently focus on evolutionarily conserved sequences surrounding the promoter of the gene of interest, which are tested for functional regulatory activity using transgenic reporter assays. This technique is effective for individual genes but will often miss elements located away from the promoter. Other methods such as ChIP-seq measure transcription factor (TF) binding genome-wide produce data that can be mined to identify candidate CRMs. However, ChIP-seq requires prior knowledge of relevant TFs and availability of ChIP-grade antibodies. Deep sequencing of DNaseI treated chromatin can be used to map individual TF DNA binding sites with high resolution and this data can be mined to identify de novo active CRMs in the genome. We are adapting DNase-seq to nuclei isolated from C. elegans. We expect to generate high-resolution maps of TF binding in the worm at different developmental stages and will use this data to discover candidate active CRMs.
Affiliation:
- Division of Biology, California Institute of Technology, Pasadena, CA.