Karlsson Christine, Larsson Jonas, Baudet Aurélie
Molecular Medicine and Gene Therapy, Lund Stem Cell Center, Lund University, Lund, Sweden.
Methods Mol Biol. 2010;650:29-43. doi: 10.1007/978-1-60761-769-3_3.
Identifying the genes and pathways that regulate self-renewal and differentiation in somatic stem cells is a central goal in stem cell and cancer biology. Here, we describe a method for RNAi-based screens in primary human hematopoietic stem and progenitor cells. These cells are suitable targets for complex, selection-based screens using pooled lentiviral shRNA libraries. The screening approach is a promising new tool to dissect regulatory mechanisms in hematopoietic and somatic stem cells, in general, and may be particularly useful to identify gene targets and modifiers that can be further exploited in strategies for ex vivo stem cell expansion.
识别调控体细胞干细胞自我更新和分化的基因及信号通路是干细胞生物学和癌症生物学的核心目标。在此，我们描述了一种在原代人类造血干细胞和祖细胞中进行基于RNA干扰（RNAi）筛选的方法。这些细胞是使用慢病毒shRNA文库进行复杂的基于选择的筛选的合适靶标。总体而言，这种筛选方法是剖析造血干细胞和体细胞干细胞调控机制的一种很有前景的新工具，对于识别可在体外干细胞扩增策略中进一步利用的基因靶标和修饰因子可能特别有用。
