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Blood, 1 August 2005, Vol. 106, No. 3, pp. 818-826. Prepublished online as a Blood First Edition Paper on April 14, 2005; DOI 10.1182/blood-2004-10-3959. This Article Full Text Full Text (PDF) All Versions of this Article: 2004-10-3959v1 106/3/818    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Lee, S.-K. Articles by Shankar, P. Search for Related Content PubMed PubMed Citation Articles by Lee, S.-K. Articles by Shankar, P. Related Collections Immunobiology Gene Therapy Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  GENE THERAPY Lentiviral delivery of short hairpin RNAs protects CD4 T cells from multiple clades and primary isolates of HIV Sang-Kyung Lee, Derek M. Dykxhoorn, Priti Kumar, Shahin Ranjbar, Erwei Song, Laura E. Maliszewski, Vanessa François-Bongarçon, Anne Goldfeld, N. Manjunath Swamy, Judy Lieberman, and Premlata Shankar From The CBR Institute for Biomedical Research and Department of Pediatrics, Harvard Medical School, Boston, MA. Viral heterogeneity is a major hurdle for potential therapeutic use of RNA interference (RNAi) against HIV-1. To determine the extent of RNAi tolerance to mutations, we tested 3 viral target sites with differing propensity for mutations: a highly variable rev sequence, a gag sequence conserved only among clade B isolates, and a vif sequence highly conserved across clades. Lentiviral expression of all 3 shRNAs inhibited replication of the homologous HIVIIIB strain. However, they differed in their ability to protect primary CD4 T cells against multiple isolates within and across HIV clades. The least conserved rev sequence inhibited only 2 of 5 clade B isolates. The gag sequence (conserved within clade B) protected 5 of 5 clade B isolates but not other clade viruses with 2 or 3 mutations in the central region. In contrast, the vif sequence, which was conserved across clades except for single mutations at positions 14 and 17, inhibited viruses from 5 different clades. Moreover, siRNAs with introduced mutations at sites of gag sequence polymorphisms showed reduced antiviral activity, whereas mutations in vif siRNA only modestly decreased silencing. Thus, although 1 or 2 mutations at peripheral sites are tolerated, mutations in the central target cleavage region abolish RNAi activity. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: E. M. Westerhout and B. Berkhout A systematic analysis of the effect of target RNA structure on RNA interference Nucleic Acids Res., July 26, 2007; 35(13): 4322 - 4330. [Abstract] [Full Text] [PDF] D. M. Dykxhoorn, L. D. Schlehuber, I. M. London, and J. Lieberman Determinants of specific RNA interference-mediated silencing of human beta-globin alleles differing by a single nucleotide polymorphism PNAS, April 11, 2006; 103(15): 5953 - 5958. [Abstract] [Full Text] [PDF] R. Sabariegos, M. Gimenez-Barcons, N. Tapia, B. Clotet, and M. A. Martinez Sequence Homology Required by Human Immunodeficiency Virus Type 1 To Escape from Short Interfering RNAs J. Virol., January 15, 2006; 80(2): 571 - 577. [Abstract] [Full Text] [PDF] S. Bartz and A. L. Jackson How Will RNAi Facilitate Drug Development? Sci. Signal., August 2, 2005; 2005(295): pe39 - pe39. [Abstract] [Full Text] [PDF]

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