High-Throughput Identification of Genes Promoting Neuron Formation and Lineage Choice in Mouse Embryonic Stem Cells

Karolinska Institute, Cell and Developmental Biology, Box 285, Stockholm 17177, Sweden.
Stem Cells (Impact Factor: 6.52). 07/2007; 25(6):1539-45. DOI: 10.1634/stemcells.2006-0485
Source: PubMed


The potential of embryonic stem cells to differentiate to all cell types makes them an attractive model for development and a potential source of cells for transplantation therapies. Candidate approaches have identified individual genes and proteins that promote the differentiation of embryonic stem cells to desired fates. Here, we describe a rapid large-scale screening strategy for the identification of genes that influence the pluripotency and differentiation of embryonic stem cells to specific fates, and we use this approach to identify genes that induce neuron formation. The power of the strategy is validated by the fact that, of the 15 genes that resulted in the largest increase in neuron number, 8 have previously been implicated in neuronal differentiation or survival, whereas 7 represent novel genes or known genes not previously implicated in neuronal development. This is a simple, fast, and generally applicable strategy for the identification of genes promoting the formation of any specific cell type from embryonic stem cells. Disclosure of potential conflicts of interest is found at the end of this article.

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Available from: Anna Falk, Nov 03, 2014
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    • "During the course of this study a report was published by Falk et al. (2007) describing a high throughput transfection screen for neuronal differentiation factors. They identified 15 genes whose expression weakly induced differentiation, none of which overlapped with ours. "
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    • "Therefore, the membrane protrusions formed in HEK293 cells may bear resemblance to polarized neurites (Shaw et al., 2002; Nalvarte et al., 2004; Govek et al., 2005). In this context it is noteworthy that a close homolog of ORP3, ORP6, was identified as one of the fifteen proteins that promote the differentiation of neurons from murine embryonic stem cells (Falk et al., 2007). Protrusion formation similar to that induced by ORP3 has been reported to occur upon overexpression of several other proteins, all of which represent regulators of the actin cytoskeleton: Cdc42 effector protein 1 (CEP1/MSE55) (Burbelo et al., 1999), ATPbinding cassette transporter-1 (ABCA1) (Wang, N. et al., 2000; Tsukamoto et al., 2001), calpain 2 (CAPN2) (Franco et al., 2004; Perrin et al., 2006), downstream of kinase 2 (Dok-2, Dok-R or FRIP) (Master et al., 2003) and c-Abl tyrosine kinase (Master et al., 2003). "
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