Project description:Dichaete is a developmentally important transcription factor, known to be involved in basic biological processes including segmentation and nervous system development among others. The aim of this experiment was to gain further insight into the role of Dichaete during early embryogenesis, specifically looking at targets in the midline using dominant negative constructs expressed via the UAS/Gal4 system, using simGal4 to drive expression in the midline.
Project description:Dichaete is a developmentally important transcription factor, known to be involved in basic biological processes including segmentation and nervous system development among others. The aim of this experiment was to gain further insight into the role of Dichaete during early embryogenesis, specifically looking at targets in the midline using dominant negative constructs expressed via the UAS/Gal4 system, using simGal4 to drive expression in the midline. 4 independent biological replicates. 3.5-4.5h old embryos were collected, and the RNA was extracted using Trizol. The UAS-dominant negative construct expressing embryos were compared to UAS-GFP expressing controls. Construct expression was driven using simgal4.
Project description:Dichaete is a developmentally important transcription factor, known to be involved in basic biological processes including segmentation and nervous system development among others. The aim of this experiment was to gain further insight into the role of Dichaete during early embryogenesis, specifically looking at targets in the midline using dominant negative constructs expressed via the UAS/Gal4 system, using prosGal4 to drive expression in developing neuroblasts.
Project description:Dichaete is a developmentally important transcription factor, known to be involved in basic biological processes including segmentation and nervous system development among others. The aim of this experiment was to gain further insight into the role of Dichaete during early embryogenesis, specifically looking at targets in the midline using dominant negative constructs expressed via the UAS/Gal4 system, using prosGal4 to drive expression in developing neuroblasts. 4 independent biological replicates. 3.5-4.5h old embryos were collected, and the RNA was extracted using Trizol. The UAS-dominant negative construct expressing embryos were compared to UAS-GFP expressing controls. Construct expression was driven using prosGal4.
Project description:Transcriptional analysis of Drosophila CNS midline cells provides a useful system for studying transcriptional regulation at the genomic level due to a well-characterized set of midline-expressed genes and in vivo-validated enhancers. To investigate how access to regulatory information is controlled during midline cell development, we performed FAIRE-seq on FACS-purified midline cells. We find that regions of the genome with a strong midline FAIRE peak and weak whole-embryo FAIRE peak (“midline-enriched”) often correspond to midline enhancers near a midline-expressed gene and provides a useful predictive tool. Analysis of a large collection of midline enhancers revealed that those with a midline-enriched peak were located near midline-expressed genes, as were about half of the midline enhancers that overlapped a midline FAIRE peak that was not statistically "midline-enriched". There is a substantial fraction of midline enhancers that do not have a midline FAIRE peak and these tend to reside in chromatin regions that are not transcriptionally active and thus, unlikely to drive midline activity in vivo. This study emphasizes the utility of combining chromatin analysis of purified cell populations with gene expression and enhancer datasets to determine an experimentally useful chromatin profile that identifies biologically relevant enhancers and genes.
Project description:Transcriptome analysis of Drosophila CNS midline cells reveals diverse peptidergic properties and a role for castor in neuronal differentiation
