Advances in Transfection Microarray^ -based studies of neuronal cell lines

Transfection Microarrays (TMA) are advantageous tools for the study of genetic events in living cells in a high-throughput functional assay. The limited numbers of cell types that can be transfected on-chip however, hinders their use. Based on our observation that ECM proteins have a significant influence on the transfection efficiency, we present herein the realization of a TMA applicable to neuronal cell lines. Further, we have conclusively proven that when siRNA was co-arrayed with the target DNA, an inhibition of neurite extension was observed. Thus, massively miniaturized RNAi experiments can now be performed in neuronal cell lines, previously unusable with transfection microarrays (TMA). Introduction Various microarray formats became more and more common in an increasingly broad spectrum of biochemical studies. The significant advantage of microarrays when compared to classical techniques is represented by low sample consumption as well as the high degree of parallelism achieved. This leads to higher throughput and therefore can allow for a comprehensive analysis of multivariant systems. The strategy for finding the genetic determinants of various processes, such as cell proliferation and differentiation, is still missing the high throughput technologies needed to collect sufficient, highly parallel and relevant gene expression data. Such high-throughput formats are expected to provide such data, on which one can base the analysis of genetic net39 Cytometry Research 14(2) : 39~44,2004 Cell operation research group Research Institute of Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST) 受付日:平成16年10月12日 受理日:平成16年11月22日 Advances in Transfection Microarray -based studies of neuronal cell lines Eiichiro Uchimura Ph.D., Shigeru Yamada Ph.D., Kayo Matsumoto, Michiko Kishi, Tomohiro Yoshikawa Ph.D., Daniel P. Funeriu Ph.D., Masato Miyake Ph.D., Jun Miyake Ph.D. Transfection Microarray技術とその応用-神経系細胞への応用 内村英一郎 ,山田 茂 ,松本圭世 ,岸 道子 ,吉川智啓 , Daniel P. Funeriu ,三宅正人 ,三宅 淳 独立行政法人産業技術総合研究所セルエンジニアリング研究部門 細胞ナノ操作工学研究グループ 抄録 トランスフェクションマイクロアレイ(TMA)は生細胞を用いた大規模網羅的な機能解析のための実験手法として 有用である。これまでは、限られた種類の細胞にのみ適用可能であったためその有用性が必ずしも生かされてはい なかった。我々はECMタンパク質を最適に用いることによりトランスフェクション効率を改善することに成功した。 それによりTMAを神経系細胞に適応することを可能とした。本報告において、これまで困難であった、神経系細胞 に対してsiRNAをもちいた特異的な機能阻害実験のマイクロアレイ化ができることを明らかとした。