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, Premlata Shankar

Research output: Contribution to journalArticle

79 Scopus citations

Abstract

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 HIV IIIB 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.

Original languageEnglish (US)
Pages (from-to)818-826
Number of pages9
JournalBlood
Volume106
Issue number3
DOIs
StatePublished - Aug 1 2005

ASJC Scopus subject areas

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology

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    Lee, S. K., Dykxhoorn, D. M., Kumar, P., Ranjbar, S., Song, E., Maliszewski, L. E., François-Bongarçon, V., Goldfeld, A., Swamy, N. M., Lieberman, J., & Shankar, P. (2005). Lentiviral delivery of short hairpin RNAs protects CD4 T cells from multiple clades and primary isolates of HIV. Blood, 106(3), 818-826. https://doi.org/10.1182/blood-2004-10-3959