Open AccessRNAi Methodologies for the Functional Study of Signaling Molecules
Author(s) -
Gwang Lee,
Leah A. Santat,
Mi Sook Chang,
Sangdun Choi
Publication year - 2009
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0004559
Subject(s) - rna interference , gene knockdown , small hairpin rna , small interfering rna , gene silencing , biology , rna silencing , microbiology and biotechnology , trans acting sirna , computational biology , cell signaling , gene , rna , signal transduction , genetics
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.
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