Yamauchi Fumio, Okada Mitsuhiro, Kato Koichi, Jakt Lars Martin, Iwata Hiroo
Institute for Frontier Medical Sciences, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
Biochim Biophys Acta. 2007 Aug;1770(8):1085-97. doi: 10.1016/j.bbagen.2007.04.005. Epub 2007 Apr 20.
Functional genomics is a central topic of current biological research, where a reverse genetic approach is one of the most promising strategies to discover functions of novel genes. Such an approach requires high-throughput methodologies to assess biological functions for a huge number of genes. We have developed a transfection array that permits parallel introduction of multiple plasmids separately into living cells. The feasibility of this array was examined in an assay system. Eleven genes were over-expressed alone, or in combination in vascular progenitors derived from embryonic stem cells. Endothelial differentiation of the cells was monitored through a stably transformed EGFP reporter construct that is expressed only in endothelial cells. Transcriptional activators that promote endothelial differentiation, such as Ets1 and Sox7, were identified. In addition, the assays also revealed an inhibitory effect on endothelial differentiation by several of the factors. These results demonstrate the feasibility of the transfection array for use in cell-based, high-throughput functional assays.
功能基因组学是当前生物学研究的核心主题，其中反向遗传学方法是发现新基因功能最具前景的策略之一。这种方法需要高通量方法来评估大量基因的生物学功能。我们开发了一种转染阵列，可将多个质粒分别平行导入活细胞。在一个检测系统中检验了该阵列的可行性。在源自胚胎干细胞的血管祖细胞中单独或组合过表达了11个基因。通过仅在内皮细胞中表达的稳定转化的EGFP报告构建体监测细胞的内皮分化。鉴定出促进内皮分化的转录激活因子，如Ets1和Sox7。此外，这些检测还揭示了几种因子对内皮分化的抑制作用。这些结果证明了转染阵列用于基于细胞的高通量功能检测的可行性。
