Abstract
RNA interference (RNAi) plays novel roles in both host antiviral defense and viral replication. It has been shown that some viruses can exploit the RNAi machinery for their own benefit by encoding for their own viral small RNAs. Here we present a collection of methods to study adenoviral small RNAs, specifically a method for immunopurification of RNA-induced silencing complex (RISC) and a biochemical assay for the activity of purified RISC associated with adenoviral small RNAs.
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References
Kumagai, M. H., Donson, J., della-Cioppa, G., Harvey, D., Hanley, K., and Grill, L. K. (1995) Cytoplasmic inhibition of carotenoid biosynthesis with virus-derived RNA. Proc Natl Acad Sci U S A. 92, 1679–1683.
Ruiz, M. T., Voinnet, O., and Baulcombe, D. C. (1998) Initiation and maintenance of virus-induced gene silencing. Plant Cell. 10, 937–946.
Cullen, B. R. (2009) Viral and cellular messenger RNA targets of viral microRNAs. Nature. 457, 421–425.
Andersson, M. G., Haasnoot, P. C., Xu, N., Berenjian, S., Berkhout, B., and Akusjärvi, G. (2005) Suppression of RNA interference by adenovirus virus-associated RNA. J Virol. 79, 9556–9565.
Aparicio, O., Razquin, N., Zaratiegui, M., Narvaiza, I., and Fortes, P. (2006) Adenovirus virus-associated RNA is processed to functional interfering RNAs involved in virus production. J Virol. 80, 1376–1384.
Sano, M., Kato, Y., and Taira, K. (2006) Sequence-specific interference by small RNAs derived from adenovirus VAI RNA. FEBS Lett. 580, 1553–1564.
Wold, W. S. M., and Tollefson, A. E. (2007) Adenovirus methods and protocols. Methods in Molecular Medicine Vol. 131. Humana, Totowa, NJ.
Meister, G., Landthaler, M., Patkaniowska, A., Dorsett, Y., Teng, G., and Tuschl, T. (2004) Human Argonaute2 mediates RNA cleavage targeted by miRNAs and siRNAs. Mol Cell. 15, 185–197.
Zamore, P. D., Tuschl, T., Sharp, P. A., and Bartel, D. P. (2000) RNAi: double-stranded RNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals. Cell. 101, 25–33.
Tuschl, T., Zamore, P. D., Lehmann, R., Bartel, D. P., and Sharp, P. A. (1999) Targeted mRNA degradation by double-stranded RNA in vitro. Genes Dev. 13, 3191–3197.
Xu, N., Segerman, B., Zhou, X., and Akusjärvi, G. (2007) Adenovirus virus-associated RNAII-derived small RNAs are efficiently incorporated into the RNA-induced silencing complex and associate with polyribosomes. J Virol. 81, 10540–10549.
Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A., and Struhl, K. (Ed.) (1995) Current Protocols in Molecular Biology. Wiley, New York.
Fritsch, E. F., Sambrook, J., Maniatis, T. (Ed.) (1989) Molecular Cloning. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Acknowledgments
This work was supported by the Swedish Cancer Society and the Uppsala RNA Research Centre (URRC).
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© 2011 Humana Press
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Xu, N., Akusjärvi, G. (2011). Characterization of RISC-Associated Adenoviral Small RNAs. In: van Rij, R. (eds) Antiviral RNAi. Methods in Molecular Biology, vol 721. Humana Press. https://doi.org/10.1007/978-1-61779-037-9_11
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DOI: https://doi.org/10.1007/978-1-61779-037-9_11
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