Project description:We performed ChIP-chip analysis using sheared chromatin isolated from Drosophila embryos of 1-3 hours in age. Keywords: high-resolution tiling array, drosophila, twist, dorsal, snail, weckle ChIP-chip of twist, dorsal, snail, and weckle
Project description:Identifying putative transcription factor target genes by combining CRISPR/Cas9-based transcriptional activation with RNAseq in Drosophila S2R+ cells. This study focuses on the transcription factors Twist and Snail, singly and together. RNA from Drosophila cells following CRISPR/Cas9-based activation of Twist, Snail, or Twist and Snail together, compared with non-targeting sgRNA. Two biological replicates for each experiment
Project description:Genetic studies have identified numerous sequence-specific transcription factors that control development, yet little is known about their in vivo distribution across animal genomes. We determine the genome-wide occupancy of the dorsoventral determinants Dorsal, Twist and Snail in the Drosophila embryo using ChIP-chip analysis. There is a tight correlation between the limits of known enhancers and the in vivo binding of these proteins. The analysis predicts substantially more target genes than previously estimated, including Dpp signaling components and genes encoding anteroposterior segmentation determinants. Thus, the ChIP-chip data uncovers a much larger network, which integrates diverse patterning processes during development.
Project description:Genetic studies have identified numerous sequence-specific transcription factors that control development, yet little is known about their in vivo distribution across animal genomes. We determine the genome-wide occupancy of the dorsoventral determinants Dorsal, Twist and Snail in the Drosophila embryo using ChIP-chip analysis. There is a tight correlation between the limits of known enhancers and the in vivo binding of these proteins. The analysis predicts substantially more target genes than previously estimated, including Dpp signaling components and genes encoding anteroposterior segmentation determinants. Thus, the ChIP-chip data uncovers a much larger network, which integrates diverse patterning processes during development.
Project description:Identifying putative transcription factor target genes by combining CRISPR/Cas9-based transcriptional activation with RNAseq in Drosophila S2R+ cells. This study focuses on the transcription factors Twist and Snail, singly and together.
Project description:Chromosome conformation capture (4C-Seq) in Drosophila Twist-H2B embryos (carrying nuclear tag specifically in the mesoderm) during embryogenesis was performed, anchoring on 107 different viewpoints. Two timepoints (3-4hrs and 6-8hrs after egg laying) and two tissue context (whole embryo and mesoderm) were assayed. Two independent collections were performed at each timepoint.
Project description:Chromosome conformation capture (4C-Seq) in Drosophila embryos from a wild-type line and from transgenic fly lines carrying the E3 enhancer of twist at ectopic locations. Two time points (2-5 hrs and 5-8 hrs after egg lay) and two viewpoints located near the twist promoter were assayed. Two independent collections were performed at each timepoint and each viewpoint.
Project description:Embryonic development requires the establishment of directional axes. In Drosophila, spatially discrete expression of transcription factors determines the anterior to posterior organization of the early embryo, while the Toll and TGF-beta signalling pathways determine the early dorsal to ventral pattern. Embryonic MAPK/ERK signaling contributes to both anterior to posterior patterning in the terminal regions and to dorsal to ventral patterning during oogenesis and embryonic stages. Here we describe a novel loss of function mutation in the Raf kinase gene, which leads to loss of ventral cell fates as seen through the loss of the ventral furrow, the absence of Dorsal/NF-kB nuclear localization, the absence of mesoderm determinants Twist and Snail, and the expansion of TGF-beta. Gene expression analysis showed cells adopting ectodermal fates similar to loss of Toll signaling. Our results combine novel mutants, live imaging, optogenetics and transcriptomics to establish a novel role for Raf, that appears to be independent of the MAPK cascade, in embryonic patterning.
