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Blood. 2009 Apr 16;113(16):3690-5. doi: 10.1182/blood-2008-10-176396. Epub 2009 Feb 2.

Forward RNAi screens in primary human hematopoietic stem/progenitor cells.

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1
Center for Regenerative Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, USA.

Abstract

The mechanisms regulating key fate decisions such as self-renewal and differentiation in hematopoietic stem and progenitor cells (HSPC) remain poorly understood. We report here a screening strategy developed to assess modulators of human hematopoiesis using a lentiviral short hairpin RNA (shRNA) library transduced into cord blood-derived stem/progenitor cells. To screen for modifiers of self-renewal/differentiation, we used the limited persistence of HSPCs under ex vivo culture conditions as a baseline for functional selection of shRNAs conferring enhanced maintenance or expansion of the stem/progenitor potential. This approach enables complex, pooled screens in large numbers of cells. Functional selection identified novel specific gene targets (exostoses 1) or shRNA constructs capable of altering human hematopoietic progenitor differentiation or stem cell expansion, respectively, thereby demonstrating the potential of this forward screening approach in primary human stem cell populations.

PMID:
19188664
PMCID:
PMC2670788
DOI:
10.1182/blood-2008-10-176396
[Indexed for MEDLINE]
Free PMC Article
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