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I-KCKT allows dissection-free RNA profiling of adult Drosophila intestinal progenitor cells

ABSTRACT: This SuperSeries is composed of the SubSeries listed below.

ORGANISM(S): Drosophila melanogaster

PROVIDER: GSE160128 | GEO | 2020/12/11

REPOSITORIES: GEO

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I-KCKT allows dissection-free RNA profiling of adult <i>Drosophila</i> intestinal progenitor cells.

Project description:The adult Drosophila intestinal epithelium is a model system for stem cell biology, but its utility is limited by current biochemical methods that lack cell type resolution. Here, we describe a new proximity-based profiling method that relies upon a GAL4 driver, termed intestinal-kickout-GAL4 (I-KCKT-GAL4), that is exclusively expressed in intestinal progenitor cells. This method uses UV crosslinked whole animal frozen powder as its starting material to immunoprecipitate the RNA cargoes of transgenic epitope-tagged RNA binding proteins driven by I-KCKT-GAL4 When applied to the general mRNA-binder, poly(A)-binding protein, the RNA profile obtained by this method identifies 98.8% of transcripts found after progenitor cell sorting, and has low background noise despite being derived from whole animal lysate. We also mapped the targets of the more selective RNA binder, Fragile X mental retardation protein (FMRP), using enhanced crosslinking and immunoprecipitation (eCLIP), and report for the first time its binding motif in Drosophila cells. This method will therefore enable the RNA profiling of wild-type and mutant intestinal progenitor cells from intact flies exposed to normal and altered environments, as well as the identification of RNA-protein interactions crucial for stem cell function.

| S-EPMC7803463 | biostudies-literature

I-KCKT allows dissection-free RNA profiling of adult Drosophila intestinal progenitor cells II

2020-12-11 | GSE160126 | GEO

I-KCKT allows dissection-free RNA profiling of adult Drosophila intestinal progenitor cells I

2020-12-11 | GSE160125 | GEO

I-KCKT allows dissection-free RNA profiling of adult Drosophila intestinal progenitor cells

Project description:I-KCKT allows dissection-free RNA profiling of adult Drosophila intestinal progenitor cells

| PRJNA671880 | ENA

I-KCKT allows dissection-free RNA profiling of adult Drosophila intestinal progenitor cells I

Project description:I-KCKT allows dissection-free RNA profiling of adult Drosophila intestinal progenitor cells I

| PRJNA672034 | ENA

I-KCKT allows dissection-free RNA profiling of adult Drosophila intestinal progenitor cells II

Project description:I-KCKT allows dissection-free RNA profiling of adult Drosophila intestinal progenitor cells II

| PRJNA672035 | ENA

zfh2 controls progenitor cell activation and differentiation in the adult Drosophila intestinal absorptive lineage.

Project description:Many tissues rely on resident stem cell population to maintain homeostasis. The balance between cell proliferation and differentiation is critical to permit tissue regeneration and prevent dysplasia, particularly following tissue damage. Thus, understanding the cellular processes and genetic programs that coordinate these processes is essential. Here, we report that the conserved transcription factor zfh2 is specifically expressed in Drosophila adult intestinal stem cell and progenitors and is a critical regulator of cell differentiation in this lineage. We show that zfh2 expression is required and sufficient to drive the activation of enteroblasts, the non-proliferative progenitors of absorptive cells. This transition is characterized by the transient formation of thin membrane protrusions, morphological changes characteristic of migratory cells and compensatory stem cell proliferation. We found that zfh2 acts in parallel to insulin signaling and upstream of the TOR growth-promoting pathway during early differentiation. Finally, maintaining zfh2 expression in late enteroblasts blocks terminal differentiation and leads to the formation of highly dysplastic lesions, defining a new late cell differentiation transition. Together, our study greatly improves our understanding of the cascade of cellular changes and regulatory steps that control differentiation in the adult fly midgut and identifies zfh2 as a major player in these processes.

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GAGA Factor maintains promoters in nucleosome-free conformation and allows promoter-proximal pausing

Project description:This SuperSeries is composed of the SubSeries listed below.

2015-04-04 | GSE58957 | GEO

Genomic dissection of conserved transcriptional regulation in intestinal epithelial cells

Project description:This SuperSeries is composed of the SubSeries listed below.

2017-05-02 | GSE94936 | GEO

Dissection of the human multipotent adult progenitor cell secretome by proteomic analysis.

Project description:Multipotent adult progenitor cells (MAPCs) are adult adherent stromal stem cells currently being assessed in acute graft versus host disease clinical trials with demonstrated immunomodulatory capabilities and the potential to ameliorate detrimental autoimmune and inflammation-related processes. Our previous studies documented that MAPCs secrete factors that play a role in regulating T-cell activity. Here we expand our studies using a proteomics approach to characterize and quantify MAPC secretome components secreted over 72 hours in vitro under steady-state conditions and in the presence of the inflammatory triggers interferon-? and lipopolysaccharide, or a tolerogenic CD74 ligand, RTL1000. MAPCs differentially responded to each of the tested stimuli, secreting molecules that regulate the biological activity of the extracellular matrix (ECM), including proteins that make up the ECM itself, proteins that regulate its construction/deconstruction, and proteins that serve to attach and detach growth factors from ECM components for redistribution upon appropriate stimulation. MAPCs secreted a wide array of proteases, some detectable in their zymogen forms. MAPCs also secreted protease inhibitors that would regulate protease activity. MAPCs secreted chemokines and cytokines that could provide molecular guidance cues to various cell types, including neutrophils, macrophages, and T cells. In addition, MAPCs secreted factors involved in maintenance of a homeostatic environment, regulating such diverse programs as innate immunity, angiogenesis/angiostasis, targeted delivery of growth factors, and the matrix-metalloprotease cascade.

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