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rnai methodologies for the functional study of signaling moleculesrnai信号分子的功能研究的方法
RNAi Methodologies for the Functional Study of
Signaling Molecules
1,2 3 3 1,4
Gwang Lee , Leah A. Santat , Mi Sook Chang , Sangdun Choi *
1 Department of Molecular Science and Technology, College of Natural Science, Ajou University, Suwon, Korea, 2 Institute for Medical Sciences, College of Natural Science,
Ajou University, Suwon, Korea, 3 Molecular Biology Laboratory, Alliance for Cellular Signaling, Division of Biology, California Institute of Technology, Pasadena, California,
United States of America, 4 Department of Biological Sciences, College of Natural Science, Ajou University, Suwon, Korea
Abstract
RNA interference (RNAi) was investigated with the aim of achieving gene silencing with diverse RNAi platforms that include
small interfering RNA (siRNA), short hairpin RNA (shRNA) and antisense oligonucleotides (ASO). Different versions of each
system were used to silence the expression of specific subunits of the heterotrimeric signal transducing G-proteins, G alpha
i2 and G beta 2, in the RAW 264.7 murine macrophage cell line. The specificity of the different RNA interference (RNAi)
platforms was assessed by DNA microarray analysis. Reliable RNAi methodologies against the genes of interest were then
developed and applied to functional studies of signaling networks. This study demonstrates a successful knockdown of
target genes and shows the potential of RNAi for use in functional studies of signaling molecules.
Citation: Lee G, Santat LA, Chang MS, Choi S (2009) RNAi Methodologies for the Functional Study of Signaling Molecules. PLoS ONE 4(2): e4559. doi:10.1371/
journal.pone.0004559
Editor: Terry Means, Massachusetts General Hospital/Harvard University, United States of America
Received January 2, 2009; Accepted January 13, 2009; Publi
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