Fabrication of cell-based arrays using micropatterned alkanethiol monolayers for the parallel silencing of specific genes by small interfering RNA.
The emergence of small interfering RNA (siRNA) opened a new opportunity to study gene functions in a genome. However, large-scale loss-of-function analyses further require cell-based high-throughput methods that allow simultaneous silencing of the huge number of genes by siRNA. In this study, we aim at fabricating the cell-based siRNA arrays that facilitate parallel introduction of multiple siRNAs into cultured mammalian cells. The siRNA arrays were prepared using surface chemical processes including the micropatterning of a self-assembled monolayer and the layer-by-layer assembly of siRNA and cationic lipid. We examined the feasibility of the siRNA array for the sequence-specific gene silencing in an array format. Furthermore, the effects of siRNA loading and culture period after transfection were studied to optimize cell-based assays on the siRNA arrays. The results obtained in this study demonstrated that our method provides the siRNA arrays with spatial specificity in gene silencing, which will serve to obtain a quantitative data set from the cell-based screens on siRNA arrays.
Fujimoto
Hiroyuki
H
Institute for Frontier Medical Sciences, Kyoto University, Shogoin, Kyoto 606-8507, Japan.
Yoshizako
Satoru
S
Kato
Koichi
K
Iwata
Hiroo
H
D001483:Base Sequence; D002460:Cell Line; D015870:Gene Expression; D020868:Gene Silencing; D006801:Humans; D046228:Microarray Analysis / Q000379:methods*; D034741:RNA, Small Interfering / Q000235:genetics*; D013438:Sulfhydryl Compounds / Q000737:chemistry*; D013997:Time Factors
D034741:RNA, Small Interfering; D013438:Sulfhydryl Compounds