Nucleic Acids Research Advance Access originally published online on March 16, 2008
Nucleic Acids Research 2008 36(9):2811-2824; doi:10.1093/nar/gkn109
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Nucleic Acids Research, 2008, Vol. 36, No. 9 2811-2824
© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
RNA |
Inhibition of HIV-1 by multiple siRNAs expressed from a single microRNA polycistron
Laboratory of Experimental Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam (CINIMA), Academic Medical Center, University of Amsterdam, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands
*To whom correspondence should be addressed. Tel: +31 20 566 4822; Fax: +31 20 691 6531; Email: b.berkhout{at}amc.uva.nl
Received January 28, 2008. Revised February 27, 2008. Accepted February 27, 2008.
RNA interference (RNAi) is a powerful approach to inhibit human immunodeficiency virus type 1 (HIV-1) replication. However, HIV-1 can escape from RNAi-mediated antiviral therapy by selection of mutations in the targeted sequence. To prevent viral escape, multiple small interfering RNAs (siRNAs) against conserved viral sequences should be combined. Ideally, these RNA inhibitors should be expressed simultaneously from a single transgene transcript. In this study, we tested a multiplex microRNA (miRNA) expression strategy by inserting multiple effective anti-HIV siRNA sequences in the miRNA polycistron mir-17-92. Individual anti-HIV miRNAs that resemble the natural miRNA structures were optimized by varying the siRNA position in the hairpin stem to obtain maximal effectiveness against luciferase reporters and HIV-1. We show that an antiviral miRNA construct can have a greater intrinsic inhibitory activity than a conventional short hairpin (shRNA) construct. When combined in a polycistron setting, the silencing activity of an individual miRNA is strongly boosted. We demonstrate that HIV-1 replication can be efficiently inhibited by simultaneous expression of four antiviral siRNAs from the polycistronic miRNA transcript. These combined results indicate that a multiplex miRNA strategy may be a promising therapeutic approach to attack escape-prone viral pathogens.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  Y. P. Liu, J. Gruber, J. Haasnoot, P. Konstantinova, and B. Berkhout RNAi-mediated inhibition of HIV-1 by targeting partially complementary viral sequences Nucleic Acids Res., October 1, 2009; 37(18): 6194 - 6204. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. L. Snyder, I. Ahmed, and L. F. Steel RNA polymerase III can drive polycistronic expression of functional interfering RNAs designed to resemble microRNAs Nucleic Acids Res., October 1, 2009; 37(19): e127 - e127. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Gurlevik, N. Woller, P. Schache, N. P. Malek, T. C. Wirth, L. Zender, M. P. Manns, S. Kubicka, and F. Kuhnel p53-dependent antiviral RNA-interference facilitates tumor-selective viral replication Nucleic Acids Res., July 1, 2009; 37(12): e84 - e84. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Ely, T. Naidoo, and P. Arbuthnot Efficient silencing of gene expression with modular trimeric Pol II expression cassettes comprising microRNA shuttles Nucleic Acids Res., July 1, 2009; 37(13): e91 - e91. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. W. Rhee, J. R. Stimers, W. Wang, and L. Pang Vascular Smooth Muscle-Specific Knockdown of the Noncardiac Form of the L-Type Calcium Channel by MicroRNA-Based Short Hairpin RNA as a Potential Antihypertensive Therapy J. Pharmacol. Exp. Ther., May 1, 2009; 329(2): 775 - 782. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. S. Weinberg and M. J.A. Wood Short non-coding RNA biology and neurodegenerative disorders: novel disease targets and therapeutics Hum. Mol. Genet., April 15, 2009; 18(R1): R27 - R39. [Abstract] [Full Text] [PDF]
|
|