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Drosha knock-down gene expression in HSPCs

ABSTRACT: Genome-wide expression differences are compared for a normal and mutant cell line. The data compares the effect of a gene mutation on genome-wide expression. empty vector (EV), shDrosha (shDr); shDr denotes hematopoietic stem-progenitor cells (HSPCs) transduced with a short-hairpin RNA targeting the miRNA-processing enzyme Drosha

ORGANISM(S): Homo sapiens

SUBMITTER: Krist Azizian

PROVIDER: E-GEOD-47403 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Global miRNA expression status of human hematopoietic stem–progenitor cells modulates myelopoiesis

Project description:We investigated how suppression of the most upstream microRNA–processing RNase, Drosha, affects the differentiation of human CD34+ hematopoietic stem–progenitor cells (HSPCs). We hypothesized that knock-down of Drosha would alter blood lineage development by modulating the expression of microRNAs. Lentiviral delivery to HSPCs of a short-hairpin targeting Drosha resulted in a viable phenotype with promotion of myeloid, and especially monocytic, maturation and suppression of apoptosis. Our results show that Drosha deficiency triggered a parallel upregulation of components of the RNAi machinery, including DGCR8, Dicer and Ago2. Deep sequencing analyses revealed global miRNA deficiency after Drosha short-hairpin treatment with relative maintenance of mature miR-223 expression. Restoration of miR-223 to normal levels after Drosha knock-down further enhanced monocytic maturation concomitant with the modulation of myeloid transcription factors that promoted monocytic differentiation. Our results support a miRNA accentuation model in which relative enhancement of miR-223 increases levels of PU.1 thereby promoting monocytic differentiation. CD34+ HSPCs were isolated from umbilical cord blood and transduced with an empty lentivector (EV) or a lentivector encoding a short-hairpin RNA targeting the pri-miRNA–processing enzyme, Drosha (shDrosha). EV and shDrosha transduced HSPCs were grown in liquid culture promoting myelopoiesis and sampled on days 0 and 7 for total RNA collection. Total RNA was size fractionated to enrich for the small RNA population and deep sequenced using ABI's SOLiD 4.0 platform.

2013-12-31 | E-GEOD-47519 | biostudies-arrayexpress

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Expression profile of control, Drosha or Dicer deleted LSKs

Project description:To determine genes regulated independently of microRNAs in early haematopoietic progenitors (LSKs) we compared the expression profiles of Drosha or Dicer deficient LSKs and control. Those genes differentially expressed between Drosha or Dicer deficient LSKs are likely regulated indepedently of microRNAs as either Drosha or Dicer deletion will lead to a complete and equivalent loss of microRNAs. LSKs were sorted from control, Drosha fl/fl of Dicer fl/fl mice.These cells were activated in vitro for 72 hours to induce total and equivalent deletion of Drosha or Dicer. RNA was extracted after 72 hours.3 repeats of the three groups were analyzed.

2015-09-10 | E-GEOD-61305 | biostudies-arrayexpress

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Project description:Rattus norvegicus Drosha, drosha ribonuclease III [Source:RGD Symbol;Acc:1307626], is expressed in 5 baseline experiment(s);

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