Forward RNAi screens in primary human hematopoietic stem/progenitor cells

被引：26

作者：

Ali, Nicole ^{[1
,2
]}

Karlsson, Christine ^{[3
]}

Aspling, Marie ^{[1
,2
]}

Hu, Guang ^{[4
,5
]}

Hacohen, Nir ^{[6
,7
]}

Scadden, David T. ^{[1
,2
,8
]}

Larsson, Jonas ^{[1
,2
,3
]}

机构：

[1] Harvard Univ, Sch Med, Massachusetts Gen Hosp, Ctr Regenerat Med, Boston, MA 02114 USA

[2] Harvard Univ, Harvard Stem Cell Inst, Cambridge, MA 02138 USA

[3] Lund Univ, Lund Stem Cell Ctr, Lund, Sweden

[4] Harvard Univ, Sch Med, Brigham & Womens Hosp, Dept Genet,Ctr Genet & Genom, Boston, MA 02114 USA

[5] Harvard Univ, Sch Med, Howard Hughes Med Inst, Boston, MA 02114 USA

[6] Broad Inst Massachusetts Inst Technol, Cambridge, MA USA

[7] Harvard Univ, Sch Med, Massachusetts Gen Hosp, Ctr Immunol & Inflammatory Dis, Cambridge, MA 02138 USA

[8] Harvard Univ, Dept Stem Cell & Regenerat Biol, Cambridge, MA 02138 USA

来源：

BLOOD | 2009年 / 113卷 / 16期

基金：

美国国家卫生研究院;

关键词：

NOD/SCID REPOPULATING CELLS; CULTURE-INITIATING CELLS; VECTOR TRANSDUCTION; GENETIC SCREEN; SECONDARY;

D O I：

10.1182/blood-2008-10-176396

中图分类号：

R5 [内科学];

学科分类号：

1002 ; 100201 ;

摘要：

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)

引用

页码：3690 / 3695

页数：6

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