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

78 Citations (Scopus)

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

Fingerprint

T-cells
Small Interfering RNA
HIV
RNA
T-Lymphocytes
Viruses
Mutation
Conserved Sequence
RNA Interference
Polymorphism
Antiviral Agents
Therapeutic Uses
HIV-1

ASJC Scopus subject areas

  • Hematology

Cite this

Lentiviral delivery of short hairpin RNAs protects CD4 T cells from multiple clades and primary isolates of HIV. / Lee, Sang Kyung; Dykxhoorn, Derek M; Kumar, Priti; Ranjbar, Shahin; Song, Erwei; Maliszewski, Laura E.; François-Bongarçon, Vanessa; Goldfeld, Anne; Swamy, N. Manjunath; Lieberman, Judy; Shankar, Premlata.

In: Blood, Vol. 106, No. 3, 01.08.2005, p. 818-826.

Research output: Contribution to journalArticle

Lee, SK, Dykxhoorn, DM, Kumar, P, Ranjbar, S, Song, E, Maliszewski, LE, François-Bongarçon, V, Goldfeld, A, Swamy, NM, Lieberman, J & Shankar, P 2005, 'Lentiviral delivery of short hairpin RNAs protects CD4 T cells from multiple clades and primary isolates of HIV', Blood, vol. 106, no. 3, pp. 818-826. https://doi.org/10.1182/blood-2004-10-3959
Lee, Sang Kyung ; Dykxhoorn, Derek M ; Kumar, Priti ; Ranjbar, Shahin ; Song, Erwei ; Maliszewski, Laura E. ; François-Bongarçon, Vanessa ; Goldfeld, Anne ; Swamy, N. Manjunath ; Lieberman, Judy ; Shankar, Premlata. / Lentiviral delivery of short hairpin RNAs protects CD4 T cells from multiple clades and primary isolates of HIV. In: Blood. 2005 ; Vol. 106, No. 3. pp. 818-826.
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