Forward RNAi screens in primary human hematopoietic stem/progenitor cells
Forward RNAi screens in primary human hematopoietic stem/progenitor cells
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DOI：
10.1182/blood-2008-10-176396
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发表时间：
2009-04-16
期刊：
影响因子：
20.3
通讯作者：
Larsson, Jonas
中科院分区：
文献类型：
--
作者：
Ali, Nicole;Karlsson, Christine;Larsson, Jonas
造血干细胞和祖细胞(HSPC)的自我更新和分化等关键命运决定的调控机制仍然知之甚少。我们在这里报道了一种筛选策略，利用慢病毒短发夹状RNA(ShRNA)文库转导到脐带血来源的干/祖细胞来评估人类造血调节因子。为了筛选自我更新/分化的修饰物，我们使用HSPC在体外培养条件下有限的持久性作为基线，以此作为功能选择的基线，以增强维持或扩大茎/祖潜力的shRNA。这种方法可以在大量单元格中实现复杂的池化屏幕。功能选择确定了能够改变人类造血祖分化或干细胞扩增的新的特定基因靶点(外突1)或shRNA构建体，从而证明了这种正向筛选方法在原代人类干细胞群体中的潜力。(血。2009；113：3690-3695)
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)