RNAi Methodologies for the Functional Study of Signaling Molecules
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.
Additional Information
© 2009 Lee et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Received: January 2, 2009; Accepted: January 13, 2009; Published: February 24, 2009. The authors would like to thank the members of The Alliance for Cellular Signaling (http://www.signaling-gateway.org/, Alliance for Cellular Signaling) for their assistance. Author Contributions: Conceived and designed the experiments: GL SC. Performed the experiments: LAS MSC. Wrote the paper: GL SC. This work was supported by a Korea Science and Engineering Foundation (KOSEF) grant funded by the Korean Government (MOST) (R01-2007-000-20533-0). This work was also partly supported by a Korea Research Foundation Grant funded by the Korean Government (MOEHRD) (KRF-2006-311-C00482) and by the Ajou University Research Fellowship of 2008. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The authors have declared that no competing interests exist.Attached Files
Published - journal.pone.0004559.PDF
Supplemental Material - journal.pone.0004559.s001.XLS
Supplemental Material - journal.pone.0004559.s002.XLS
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Additional details
- PMCID
- PMC2641016
- Eprint ID
- 101340
- Resolver ID
- CaltechAUTHORS:20200218-151248762
- R01-2007-000-20533-0
- Korea Science and Engineering Foundation
- KRF-2006-311-C00482
- Korea Research Foundation
- Ajou University
- Created
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2020-02-19Created from EPrint's datestamp field
- Updated
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2021-11-16Created from EPrint's last_modified field