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Forward RNAi screens in primary human hematopoietic stem/progenitor cells

Ali, Nicole ; Karlsson, Christine LU ; Aspling, Marie ; Hu, Guang ; Hacohen, Nir ; Scadden, David T. and Larsson, Jonas LU (2009) In Blood 113(16). p.3690-3695
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... (More)
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. (Blood. 2009; 113: 3690-3695) (Less)
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author
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organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Blood
volume
113
issue
16
pages
3690 - 3695
publisher
American Society of Hematology
external identifiers
  • wos:000265445900009
  • scopus:65549123755
ISSN
1528-0020
DOI
10.1182/blood-2008-10-176396
language
English
LU publication?
yes
id
cd2dd0f9-63eb-4222-8c61-d665cd8ba811 (old id 1399585)
date added to LUP
2016-04-01 12:34:45
date last changed
2022-01-27 07:00:46
@article{cd2dd0f9-63eb-4222-8c61-d665cd8ba811,
  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. (Blood. 2009; 113: 3690-3695)}},
  author       = {{Ali, Nicole and Karlsson, Christine and Aspling, Marie and Hu, Guang and Hacohen, Nir and Scadden, David T. and Larsson, Jonas}},
  issn         = {{1528-0020}},
  language     = {{eng}},
  number       = {{16}},
  pages        = {{3690--3695}},
  publisher    = {{American Society of Hematology}},
  series       = {{Blood}},
  title        = {{Forward RNAi screens in primary human hematopoietic stem/progenitor cells}},
  url          = {{http://dx.doi.org/10.1182/blood-2008-10-176396}},
  doi          = {{10.1182/blood-2008-10-176396}},
  volume       = {{113}},
  year         = {{2009}},
}