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Polymerase II binding analysis during nutrient restriction using targeted DamID

ABSTRACT: Polymerase II binding analysis during nutrient restriction using targeted DamID

PROVIDER: PRJNA605807 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR11058559.fastq.gz	Fastqsanger.gz
	SRR11058560.fastq.gz	Fastqsanger.gz
	SRR11058561.fastq.gz	Fastqsanger.gz
	SRR11058562.fastq.gz	Fastqsanger.gz
	SRR11058563.fastq.gz	Fastqsanger.gz

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Polymerase II binding analysis during nutrient restriction using targeted DamID

Project description:The adaptable transcriptional response to changes in food availability not only ensures animal survival, but also permits progressing with embryonic development. Interestingly, the central nervous system is preferentially protected to periods of malnutrition, a phenomenon known as ‘brain sparing’. However, the mechanisms that mediates this genetic response remains poorly understood. To get a better understanding of this, we used Drosophila melanogaster as a genetic model, analysing the transcriptional response of larval neural stem cells (neuroblasts) and glial cells of the blood-brain barrier during nutrient restriction using the targeted DamID technique.

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Project description:To adapt RNA polymerase DamID (RAPID) for FLP-mediated spatial control in Caenorhabditis elegans, we inserted a Dam::rpb-6 fusion gene downstream of a FRT-flanked mCh::his-58 cassette and under control of the hsp-16.41 promoter. We introduced a single copy of this construct into the C. elegans genome and crossed the resulting line with a dpy-7p::FLP driver to enable basal Dam::rpb-6 expression in the hypodermis. Nematodes were cultured at 20 degrees Celcius to ensure low levels of Dam::RPB-6 expression in the hypodermis and total genomic DNA was purified from L4 larvae. DNA from animals expressing GFP::Dam was used to control for unspecific methylation. The genome-wide association profile of Dam::RPB-6 was determined by deep sequencing, which revealed a list of 2331 protein coding genes with FDR < 0.05. Original RAPID reference: Gomez-Saldivar et al (2020) Tissue-Specific Transcription Footprinting Using RNA PoI DamID (RAPID) in Caenorhabditis elegans. Genetics 216, 931–945. doi:10.1534/genetics.120.303774

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