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MiR-150 Pulldown-sequencing data

ABSTRACT: miR-150 Pulldown-sequencing data

PROVIDER: PRJNA675569 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13010521.fastq.gz	Fastqsanger.gz
	SRR13010522.fastq.gz	Fastqsanger.gz
	SRR13010523.fastq.gz	Fastqsanger.gz
	SRR13010524.fastq.gz	Fastqsanger.gz
	SRR13010525.fastq.gz	Fastqsanger.gz

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miR-150 Pulldown-sequencing data

Project description:Human CD4+ Total memory cells (CD4+CD25-CD45RA-CCR7+/-) were activated for 5d with anti-CD3/anti-CD28 antibodies and transfected with biotinylated miR-150 mimic or biotinylated mimic negative control by Exiqon. After 24h cells were lysed and miRNA:mRNA complexes pulled-down with streptavidin agarose O/N at 4°C. RNA was then purified using TRI-reagent and RNA zymospin columns. Three independent donors were used for this experiment. Inputs were also sequenced. Total RNA was sequenced in single end mode. This experiment is assessing direct targets bound by miR-150-5p in human T cells.

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Project description:Here we show that biotin-labelled miR-34a can be loaded to AGO2, and AGO2 immunoprecipitation can pulldown biotinylated miR-34a (Bio-miR pulldown). RNA-sequencing (RNA-seq) of the Bio-miR pulldown RNAs efficiently identified miR-34a mRNA targets, which could be verified with luciferase assays. In contrast to the approach of Bio-miR pulldown, RNA-seq of miR-34a overexpression samples had limited value in identifying direct targets of miR-34a. It seems that pulldown of 30 -Biotin-tagged miRNA can identify bona fide microRNA targets at least for miR34a.

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CTCL and ATL cell lines, mir-150 transfected versus controls

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miR-146a Pulldown-sequencing data

Project description:Human CD4+ Total memory cells (CD4+CD25-CD45RA-CCR7+/-) were activated with anti-CD3/anti-CD28 antibodies and transfected with biotinylated miR-146a mimic or biotinylated mimic negative control by Exiqon. After 24h cells were lysed and miRNA:mRNA complexes pulled-down with streptavidin agarose O/N at 4°C. RNA was then purified using TRI-reagent and RNA zymospin columns. Four independent donors were used for this experiment. Total RNA was sequenced in 2 lanes, paired ends. This experiment is assessing direct targets bound by miR-146a in human T cells.

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Project description:MicroRNAs are a major class of gene regulators in mammals. While numerous aspects of the immune systems are controlled by miRNAs, their precise role in the CD8+ T cell response remains unclear. In this report, we show that miR-150 is the most abundant miRNA expressed in CD8+ T cells and its expression is required for proper effector cell differentiation in response to acute and chronic pathogens. In the absence of miR-150, CD8+ T cells failed to both undergo robust expansion and differentiate into short-lived terminal effector cells. The lack of miR-150 also altered the effector CD8+ T cell transcriptome such that, despite activation, genes associated with naïve or memory cells were highly expressed. The deletion of miR-150 also reduced killing efficiency of CD8+ T cells. These results uncover a cell-intrinsic role for miR-150 in the regulation of CD8+ T cell effector fate and function.

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