FoxD-a/b ChIP-chip on Ciona intestinalis embryo (32-cell stage)
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ABSTRACT: A genomic overview of in vivo binding of a transcription factor FoxD-a/b in the ascidian 32-cell stage embryo. Fertilized eggs were electroporated with a construct encoding GFP-tagged FoxD-a/b. After fixation, chromatin was sonicated to obtain DNA fragments with an average of size of 1kb. The DNA fragments were enriched by immunoprecipitation with an polyclonal antibody specific for GFP (Invitrogen). The immunoprecipitated and WCE (Whole Cell Extract) DNA were amplified by LM-PCR for the Chip analysis. Two independent experiments were performed to reduce experimental errors.
Project description:A genomic overview of in vivo binding of a transcription factor Snail in the ascidian early gastrula embryo. Fertilized eggs were electroporated with a construct encoding GFP-tagged Snail. After fixation, chromatin was sonicated to obtain DNA fragments with an average of size of 1kb. The DNA fragments were enriched by immunoprecipitation with an polyclonal antibody specific for GFP (Invitrogen). The immunoprecipitated and WCE (Whole Cell Extract) DNA were amplified by LM-PCR for the Chip analysis. Two independent experiments were performed to reduce experimental errors.
Project description:A genomic overview of in vivo binding of a transcription factor Neurogenin in the ascidian 64-cell stage embryo. Fertilized eggs were electroporated with a construct encoding GFP-tagged Neurogenin. After fixation, chromatin was sonicated to obtain DNA fragments with an average of size of 1kb. The DNA fragments were enriched by immunoprecipitation with an polyclonal antibody specific for GFP (Invitrogen). The immunoprecipitated and WCE (Whole Cell Extract) DNA were amplified by LM-PCR for the Chip analysis. Two independent experiments were performed to reduce experimental errors.
Project description:A genomic overview of in vivo binding of a transcription factor Tbx6b in the ascidian early gastrula embryo. Fertilized eggs were electroporated with a construct encoding GFP-tagged Tbx6b. After fixation, chromatin was sonicated to obtain DNA fragments with an average of size of 1kb. The DNA fragments were enriched by immunoprecipitation with an polyclonal antibody specific for GFP (Invitrogen). The immunoprecipitated and WCE (Whole Cell Extract) DNA were amplified by LM-PCR for the Chip analysis. Two independent experiments were performed to reduce experimental errors.
Project description:A genomic overview of in vivo binding of a transcription factor Otx in the ascidian early gastrula embryo. Fertilized eggs were electroporated with a construct encoding GFP-tagged Otx. After fixation, chromatin was sonicated to obtain DNA fragments with an average of size of 1kb. The DNA fragments were enriched by immunoprecipitation with an polyclonal antibody specific for GFP (Invitrogen). The immunoprecipitated and WCE (Whole Cell Extract) DNA were amplified by LM-PCR for the Chip analysis. Two independent experiments were performed to reduce experimental errors.
Project description:A genomic overview of in vivo binding of a transcription factor SoxC in the ascidian early gastrula embryo. Fertilized eggs were electroporated with a construct encoding GFP-tagged SoxC. After fixation, chromatin was sonicated to obtain DNA fragments with an average of size of 1kb. The DNA fragments were enriched by immunoprecipitation with an polyclonal antibody specific for GFP (Invitrogen). The immunoprecipitated and WCE (Whole Cell Extract) DNA were amplified by LM-PCR for the Chip analysis. Two independent experiments were performed to reduce experimental errors.
Project description:A genomic overview of in vivo binding of a transcription factor Twist-like-1a/b in the ascidian early gastrula embryo. Fertilized eggs were electroporated with a construct encoding GFP-tagged Twist-like-1a/b. After fixation, chromatin was sonicated to obtain DNA fragments with an average of size of 1kb. The DNA fragments were enriched by immunoprecipitation with an polyclonal antibody specific for GFP (Invitrogen). The immunoprecipitated and WCE (Whole Cell Extract) DNA were amplified by LM-PCR for the Chip analysis. Three independent experiments were performed to reduce experimental errors.
Project description:A genomic overview of in vivo binding of a transcription factor FoxD-a/b in the ascidian 64-cell stage embryo. Fertilized eggs were electroporated with a construct encoding GFP-tagged FoxD-a/b. After fixation, chromatin was sonicated to obtain DNA fragments with an average of size of 1kb. The DNA fragments were enriched by immunoprecipitation with an polyclonal antibody specific for GFP (Invitrogen). The immunoprecipitated and WCE (Whole Cell Extract) DNA were amplified by LM-PCR for the Chip analysis. Two independent experiments were performed to reduce experimental errors.
Project description:A genomic overview of in vivo binding of a transcription factor FoxA-a in the ascidian 64-cell stage embryo. Fertilized eggs were electroporated with a construct encoding GFP-tagged FoxA-a. After fixation, Chromatin was sonicated to obtain DNA fragments with an average of size of 1kb. The DNA fragments were enriched by immunoprecipitation with an polyclonal antibody specific for GFP (Invitrogen). The immunoprecipitated and WCE (Whole Cell Extract) DNA were amplified by LM-PCR for the Chip analysis. Two independent experiments were performed to reduce experimental errors.
Project description:A comprehensive search for downstream genes is necessary for understanding of functions of Nodal signaling in embryos of the ascidian Ciona intestinalis. The level of mRNA expression in Nodal-overexpressing embryos was compared with that in normal embryos at the late gastrula stage by a microarray analysis. Overexpression of Ci-nodal elevated the level of expression of 115 genes more than two-fold. These candidate target genes include those encoding transcription factors, proteins involved in cell-cell communication, and extracellular matrix components. Many of these genes were expressed in the neural plate at the late gastrula stage. Overexpression of nodal also affected the expression of genes involved in the planar cell polarity pathway and Wnt/Ca2+ pathway (prickle, Rho-like2, and PLC2), and delta1-protocadherin-like. Keywords: ascidian, Ciona intestinalis, Nodal, embryogenesis, gene expression profile two samples Dye Swap design with 2 arrays
Project description:The transcriptome of unfertilized eggs of Ciona intestinalis was determined with microarrays. To obtain gene expression profiles of unfertilized eggs, we took 14 biological replicates. For the first through fourth replicates, we also took a technical duplicate. Gene expression profiles of the 8-, 16-, 32-cell embryos were obtained with embryos from the third to fifth animals.