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| Status |
Public on Apr 02, 2021 |
| Title |
RNAseq of Polycomb KD ISCs |
| Organism |
Drosophila melanogaster |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Tissue homeostasis requires long-term lineage fidelity of somatic stem cells. Whether and how age-related changes in somatic stem cells impact the faithful execution of lineage decisions remains largely unknown. Here, we address this question using genome-wide chromatin accessibility and transcriptome analysis as well as single cell RNA-seq to explore stem cell intrinsic changes in the aging Drosophila intestine. These studies indicate that in stem cells of old flies, promoters of Polycomb (Pc) target genes become differentially accessible, resulting in the increased expression of enteroendocrine (EE) cell specification genes. Consistently, we find age related changes in the composition of the EE progenitor cell population in aging intestines, as well as a significant increase in the proportion of EE-specified ISCs and progenitors in aging flies. We further confirm that Pc-mediated chromatin regulation is a critical determinant of EE cell specification in the Drosophila intestine. Pc is required to maintain expression of stem cell genes while ensuring repression of differentiation and specification genes. Our results identify Pc group proteins as central regulators of lineage identity in the intestinal epithelium and highlight the impact of age-related decline in chromatin regulation on tissue homeostasis.
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| Overall design |
For polycomb (Pc)-RNAi experiments, virgins of the ISCts fly line were crossed with cherry-RNAi or Pc-RNAi (BL36070) males and maintained at 18 °C. Progeny was collected and shifted to 29°C 4-7 days after eclosion. Flies were kept at 29 °C for 8-11 days at which point they were dissected and ISCs were isolated. In short, midguts were dissected in 1xPBS, 1% Bovine Serum Albumin (BSA) and dissociated in 0.5% Trypsin-EDTA solution for less than 2h at room temperature (RT), during which dissociated cells were collected periodically every 20-30min, re537 suspended in 1xPBS, 1%BSA and 2%FBS and kept on ice until sorting. A BD Biosciences FACSAria II flow cytometer cell sorter was used for cell sorting.
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| Contributor(s) |
Tauc H, Rodriguez-Fernandez I, Hackney J, Pawlak M, Ronnen Oron T, Korzelius J, F. Moussa H, Chaudhuri S, Modrusan Z, A. Edgar B, Jasper H |
| Citation(s) |
33724181 |
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| Submission date |
Sep 10, 2020 |
| Last update date |
Apr 03, 2021 |
| Contact name |
Michal Pawlak |
| E-mail(s) |
mpawlak@get2omics.com
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| Organization name |
Roche Polska Sp. z o.o.
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| Street address |
Domaniewska 39 B
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| City |
Warsaw |
| ZIP/Postal code |
02-672 |
| Country |
Poland |
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| Platforms (1) |
| GPL17275 |
Illumina HiSeq 2500 (Drosophila melanogaster) |
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| Samples (6)
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| GSM4775093 |
SAM24366244 2. ISCts_CherryRNAi 6.20.2019_10d Intestine Small SAM24366244 2. ISCts_CherryRNAi 6.20.2019_10d B2 |
| GSM4775094 |
SAM24366247 2. ISCts_PcRNAi_36070_ 6.20.2019_10d Intestine Small SAM24366247 2. ISCts_PcRNAi_36070_ 6.20.2019_10d B2 |
| GSM4775095 |
SAM24366246 1. ISCts_PcRNAi_36070_ 6.17.2019_10d Intestine Small SAM24366246 1. ISCts_PcRNAi_36070_ 6.17.2019_10d B1 |
| GSM4775096 |
SAM24366248 3. ISCts_PcRNAi_36070_ 6.26.2019_9d Intestine Small SAM24366248 3. ISCts_PcRNAi_36070_ 6.26.2019_9d B3 |
| GSM4775097 |
SAM24366245 3. ISCts_CherryRNAi 6.26.2019_9d Intestine Small SAM24366245 3. ISCts_CherryRNAi 6.26.2019_9d B3 |
| GSM4775098 |
SAM24366243 1. ISCts_CherryRNAi 6.17.2019_10d Intestine Small SAM24366243 1. ISCts_CherryRNAi 6.17.2019_10d B1 |
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| This SubSeries is part of SuperSeries: |
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| Relations |
| BioProject |
PRJNA662795 |
| SRA |
SRP281998 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE157793_RAW.tar |
1020.0 Kb |
(http)(custom) |
TAR (of TSV) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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