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Homo sapiens

ABSTRACT: Human Adult Sorted Live Cell Erythroblasts transduced with Sigma non-targeting shRNA negative control (SHC002V) with puromycin selection RNAseq

PROVIDER: PRJNA369084 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR5209670_1.fastq.gz	Fastqsanger.gz
	SRR5209670_2.fastq.gz	Fastqsanger.gz
	SRR5209680_1.fastq.gz	Fastqsanger.gz
	SRR5209680_2.fastq.gz	Fastqsanger.gz
	SRR5209690_1.fastq.gz	Fastqsanger.gz

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Homo sapiens

Project description:Human Adult Sorted Live Cell Erythroblasts transduced with Sigma shRNA Clone TRCN0000005418 targeting RIOK3 with puromycin selection RNAseq

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Project description:Human Adult Sorted Live Cell Erythroblasts transduced with Sigma shRNA Clone TRCN0000162889 targeting SLC25A39 with puromycin selection RNAseq.

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Human Adult Sorted Live Cell Erythroblasts transduced with Sigma non-targeting shRNA negative control (SHC002V) with puromycin selection RNAseq

Project description:Erythroblasts cultured from mobilized CD34+ cells from six healthy adult human donors were used to generate an erythroblast transcriptome transduced with Sigma non-targeting shRNA negative control . Cellular maturation was maintained including enucleation. On culture day 14 total RNA was isolated (see PMID: 23798711 for details). These RNA-Seq profiles were generated after flow cytometric sorting (live cell gating of culture Day 14 erythroblasts according to forward and side scatter). We would like to thank Gerard Bouffard and the National Human Genome Research Institute for their expertise and assistance with RNA-seq.

2017-02-01 | GSE94147 | GEO

Human Adult Sorted Live Cell Erythroblasts transduced with Sigma shRNA Clone TRCN0000005418 targeting RIOK3 with puromycin selection RNAseq

Project description:Erythroblasts cultured from mobilized CD34+ cells from healthy adult human donors were used to generate an erythroblast transcriptome transduced with Sigma shRNA Clone TRCN0000005418 targeting RIOK3 gene. Cellular maturation was maintained including enucleation. On culture day 14 total RNA was isolated (see PMID: 23798711 for details). These RNA-Seq profiles were generated after flow cytometric sorting (live cell gating of culture Day 14 erythroblasts according to forward and side scatter). Preliminary screening data demonstrated a robust increase in fetal hemoglobin that may be due to off-target effects from this shRNA clone. A non-targeting shRNA dataset was used for control. These samples were originally published in GSE94147 and are included in this series as well.

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Human Adult Sorted Live Cell Erythroblasts transduced with Sigma shRNA Clone TRCN0000162889 targeting SLC25A39 with puromycin selection RNAseq.

Project description:Erythroblasts cultured from mobilized CD34+ cells from six healthy adult human donors were used to generate an erythroblast transcriptome transduced with Sigma shRNA Clone TRCN0000162889 targeting SLC25A39 gene. Cellular maturation was maintained including enucleation. On culture day 14 total RNA was isolated (see PMID: 23798711 for details). These RNA-Seq profiles were generated after flow cytometric sorting (live cell gating of culture Day 14 erythroblasts according to forward and side scatter). Preliminary screening data demonstrated a robust increase in fetal hemoglobin that may be due to off-target effects from this shRNA clone. See Series GSE94147 for Empty Vector Control dataset.

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Project description:The 22Rv1 and PC-3 cells were transduced with shMIMIC human lentiviral vectors (Horizon Discovery, Cambridge, UK) to stably overexpress miRNA-23c or -4328. The SMARTvector Non-Targeting Control (NTC) was expressed to serve as a negative control. The vectors contained a turbo green fluorescent protein (turboGFP) and a puromycin resistance gene cassette. After transduction, cells were cultured in a medium supplemented with 5 µg/mL puromycin (Takara Bio, Tokyo, Japan) for antibiotic selection. Overexpression was confirmed by monitoring the turboGFP by fluorescence microscopy and by RT-qPCR analysis of miRNA-23c and -4328 levels. Relative protein quantification was performed to compare protein expression in 22Rv1 and PC-3 single cell clones overexpressing miRNA-23c and -4328, compared to corresponding NTC cells. Single cell clones overexpressing either miRNA-23c (n = 3), -4328 (n = 3) or NTC (n = 3) were analyzed in triplicate.

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