Abstract
RNA interference (RNAi) has attracted a lot of interest during recent years as a method to knock-down gene expression and as a possible antiviral system. Here we present a collection of in vitro methods to study RNAi and the effect of an adenovirus infection on RNAi. We describe methods to measure the two key enzymatic complexes involved in RNAi: Dicer and RISC.
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Fire, A., Xu, S., Montgomery, M. K., Kostas, S. A., Driver, S. E., andMello, C. C. (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391, 806–811.
Hutvagner, G. and Zamore, P. D. (2002) RNAi: nature abhors a double-strand. Curr. Opin. Genet. Dev. 12, 225–232.
Denli, A. M. and Hannon, G. J. (2003) RNAi: an ever-growing puzzle. Trends Biochem. Sci. 28, 196–201.
Baulcombe, D. (2002) Viral suppression of systemic silencing. Trends Microbiol. 10, 306–308.
Lecellier, C. H. and Voinnet, O. (2004) RNA silencing: no mercy for viruses? Immunol. Rev. 198, 285–303.
Li, W. X. and Ding, S. W. (2001) Viral suppressors of RNA silencing. Curr. Opin. Biotechnol. 12, 150–154.
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.
Lu, S. and Cullen, B. R. (2004) Adenovirus VA1 noncoding RNA can inhibit small interfering RNA and MicroRNA biogenesis. J. Virol. 78, 12,868–12,876.
Kreivi, J. P., Trinkle-Mulcahy, L., Lyon, C. E., Morrice, N. A., Cohen, P., and Lamond, A. I. (1997) Purification and characterisation of p99, a nuclear modulator of protein phosphatase 1 activity. FEBS Lett.420, 57–62.
Zamore, P. D., Tuschl, T., Sharp, P. A., and Bartel, D. P. (2000) RNAi: doublestranded 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.
Schwarz, D. S., Hutvagner, G., Du, T., Xu, Z., Aronin, N., and Zamore, P. D. (2003) Asymmetry in the assembly of the RNAi enzyme complex. Cell 115, 199–208.
Provost,P., Dishart, D., Doucet, J.,Frendewey, D., Samuelsson,B., and Radmark, O. (2002) Ribonuclease activity and RNA binding of recombinant human Dicer. EMBO J. 21, 5864–5874.
Rotondo, G. and Frendewey, D. (2001) Pac1 ribonuclease of Schizosaccharomyces pombe. Methods Enzymol. 342, 168–193.
Bhat, R. A. and Thimmappaya, B. (1984) Adenovirus mutants with DNA sequence perturbations in the intragenic promoter of VAI RNA gene allow the enhanced transcription of VAII RNA gene in HeLa cells. Nucleic Acids Res. 12, 7377–7388.
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© 2007 Humana Press Inc, Totowa, NJ
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Andersson, G., Xu, N., Akusjärvi, G. (2007). In Vitro Methods to Study RNA Interference During an Adenovirus Infection. In: Wold, W.S.M., Tollefson, A.E. (eds) Adenovirus Methods and Protocols. Methods in Molecular Medicine™, vol 131. Humana Press. https://doi.org/10.1007/978-1-59745-277-9_4
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DOI: https://doi.org/10.1007/978-1-59745-277-9_4
Publisher Name: Humana Press
Print ISBN: 978-1-58829-901-7
Online ISBN: 978-1-59745-277-9
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