Abstract
MicroRNAs (miRNAs) are a class of small ∼22 nt regulatory RNAs that modulate mRNA expression in all multicellular eukaryotic organisms. Interestingly, viruses also encode miRNAs and these viral miRNAs target cellular and viral mRNAs to regulate virus replication and latent infection. In particular, herpesviruses encode a large number of miRNAs. Herpesvirus infection also changes the normal expression profile of cellular miRNAs. New genetic tools have recently been generated to study the function of viral and cellular miRNAs in virus-infected cells. The creation of these reagents and use in Epstein-Barr virus-infected lymphoblastoid cell lines are discussed as a model viral system for the investigation of miRNA function.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Skalsky RL, Cullen BR (2010) Viruses, microRNAs, and host interactions. Annu Rev Microbiol 64:123–141
Ebert MS, Neilson JR, Sharp PA (2007) MicroRNA sponges: competitive inhibitors of small RNAs in mammalian cells. Nat Methods 4(9):721–726
Ebert MS, Sharp PA (2010) MicroRNA sponges: progress and possibilities. RNA 16(11):2043–2050
Soutschek J et al (2004) Therapeutic silencing of an endogenous gene by systemic administration of modified siRNAs. Nature 432(7014):173–178
Linnstaedt SD et al (2010) Virally induced cellular miR-155 plays a key role in B-cell immortalization by EBV. J Virol 84(22):11670–11678
Gottwein E, Cai X, Cullen BR (2006) A novel assay for viral microRNA function identifies a single nucleotide polymorphism that affects Drosha processing. J Virol 80(11):5321–5326
Dull T et al (1998) A third-generation lentivirus vector with a conditional packaging system. J Virol 72(11):8463–8471
Gottwein E, Cullen BR (2012) A human herpesvirus microRNA inhibits p21 expression and attenuates p21-mediated cell cycle arrest. J Virol 86(12):6889–6898. Epub 2012 Apr 11
Ameres SL et al (2011) Target RNA-directed tailing and trimming purifies the sorting of endo-siRNAs between the two Drosophila Argonaute proteins. RNA 17(1):54–63
Gottwein E et al (2007) A viral microRNA functions as an orthologue of cellular miR-155. Nature 450(7172):1096–1099
Gottwein E, Cullen BR (2007) Protocols for expression and functional analysis of viral microRNAs. Methods Enzymol 427:229–243
J Virol. 2012 Jun;86 (12); 6889-98, Epub 2112 Apr 11 3
Acknowledgments
Eva Gottwein initially developed many of these protocols in Bryan Cullen’s laboratory. Dr. Gottwein graciously shared this information in collaboration with our group to aid in the EBV-specific development of the protocols. We thank Raul Salinas for assisting in the preparation of the manuscript. This work was supported by pilot grants to M.A.L. from the Stewart Trust, the American Cancer Society, and the Duke Cancer Institute, as well as an NCI-supported pilot award for collaborations between CFARs and Cancer Centers (iCHARM) (awarded from Penn CFAR; P30-AI-045008).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Forte, E., Luftig, M.A. (2013). Use of Viral Systems to Study miRNA-Mediated Regulation of Gene Expression in Human Cells. In: Ying, SY. (eds) MicroRNA Protocols. Methods in Molecular Biology, vol 936. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-083-0_12
Download citation
DOI: https://doi.org/10.1007/978-1-62703-083-0_12
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-082-3
Online ISBN: 978-1-62703-083-0
eBook Packages: Springer Protocols