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Cell Rep. 2012 Aug 30;2(2):407-18. doi: 10.1016/j.celrep.2012.07.008. Epub 2012 Aug 9.

FACS purification and transcriptome analysis of drosophila neural stem cells reveals a role for Klumpfuss in self-renewal.

Author information

1
Institute of Molecular Biotechnology of the Austrian Academy of Science, Vienna, Austria.

Abstract

Drosophila neuroblasts (NBs) have emerged as a model for stem cell biology that is ideal for genetic analysis but is limited by the lack of cell-type-specific gene expression data. Here, we describe a method for isolating large numbers of pure NBs and differentiating neurons that retain both cell-cycle and lineage characteristics. We determine transcriptional profiles by mRNA sequencing and identify 28 predicted NB-specific transcription factors that can be arranged in a network containing hubs for Notch signaling, growth control, and chromatin regulation. Overexpression and RNA interference for these factors identify Klumpfuss as a regulator of self-renewal. We show that loss of Klumpfuss function causes premature differentiation and that overexpression results in the formation of transplantable brain tumors. Our data represent a valuable resource for investigating Drosophila developmental neurobiology, and the described method can be applied to other invertebrate stem cell lineages as well.

PMID:
22884370
PMCID:
PMC3828055
DOI:
10.1016/j.celrep.2012.07.008
[Indexed for MEDLINE]
Free PMC Article

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