Abstract
RNA interference is a potent gene silencing pathway initiated by short molecules of double-stranded RNA. Small interfering RNAs (siRNAs) with full sequence complementarity to mRNAs induce cleavage of their target transcripts in the cytoplasm. Recent evidence has shown, however, that siRNAs can also function in the nucleus of mammalian cells to affect changes in chromatin structure. When targeted to promoter regions, siRNAs load into the effector protein Argonaute-1 (AGO1) and direct the formation of silent chromatin domains. This mechanism is known as transcriptional gene silencing (TGS), and the development of TGS as a novel therapeutic modality would be applicable to chronic diseases where long-term, heritable silencing of target genes is warranted. Here we discuss how small RNAs can be used to direct TGS in mammalian cells.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Kim, D. H. and Rossi, J. J. (2007) Strategies for silencing human disease using RNA interference. Nat. Rev. Genet. 8, 173–184.
Kim, D. H., Villeneuve, L. M., Morris, K. V., and Rossi, J. J. (2006) Argonaute-1 directs siRNA-mediated transcriptional gene silencing in human cells. Nat. Struct. Mol. Biol. 13, 793–797.
Weinberg, M. S., Villeneuve, L. M., Ehsani, A., Amarzguioui, M., Aagaard, L., Chen, Z. X., Riggs, A. D., Rossi, J. J., and Morris, K. V. (2006) The antisense strand of small interfering RNAs directs histone methylation and transcriptional gene silencing in human cells. RNA 12, 256–262.
Ting, A. H., Schuebel, K. E., Herman, J. G., and Baylin, S. B. (2005) Short double-stranded RNA induces transcriptional gene silencing in human cancer cells in the absence of DNA methylation. Nat. Genet. 37, 906–910.
Janowski, B. A., Huffman, K. E., Schwartz, J. C., Ram, R., Hardy, D., Shames, D. S., Minna, J. D., and Corey, D. R. (2005) Inhibiting gene expression at transcription start sites in chromosomal DNA with antigene RNAs. Nat. Chem. Biol. 1, 216–222.
Pulukuri, S. M. and Rao, J. S. (2007) Small interfering RNA directed reversal of urokinase plasminogen activator demethylation inhibits prostate tumor growth and metastasis. Cancer Res. 67, 6637–6646.
Han, J., Kim, D., and Morris, K. V. (2007) Promoter-associated RNA is required for RNA-directed transcriptional gene silencing in human cells. Proc. Natl. Acad. Sci. USA 104, 12422–12427.
Zhang, M. X., et al. (2005) Regulation of endothelial nitric oxide synthase by small RNA. Proc. Natl. Acad. Sci. USA 102, 16967–16972.
Acknowledgments
This work was supported by a City of Hope Graduate School Internal Fellowship to D.H.K. and National Institutes of Health grants R37-AI29329 and R01-AI42552 to J.J.R.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Kim, D.H., Rossi, J.J. (2009). Transcriptional Gene Silencing Using Small RNAs. In: Rondinone, C., Reidhaar-Olson, J. (eds) Therapeutic Applications of RNAi. Methods in Molecular Biology™, vol 555. Humana Press. https://doi.org/10.1007/978-1-60327-295-7_9
Download citation
DOI: https://doi.org/10.1007/978-1-60327-295-7_9
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-60327-294-0
Online ISBN: 978-1-60327-295-7
eBook Packages: Springer Protocols