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 language | English |
|---|---|
| Pages (from-to) | 818-826 |
| Number of pages | 9 |
| Journal | Blood |
| Volume | 106 |
| Issue number | 3 |
| DOIs | |
| State | Published - Aug 1 2005 |
| Externally published | Yes |
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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 journal › Article
}
TY - JOUR
T1 - Lentiviral delivery of short hairpin RNAs protects CD4 T cells from multiple clades and primary isolates of HIV
AU - Lee, Sang Kyung
AU - Dykxhoorn, Derek M
AU - Kumar, Priti
AU - Ranjbar, Shahin
AU - Song, Erwei
AU - Maliszewski, Laura E.
AU - François-Bongarçon, Vanessa
AU - Goldfeld, Anne
AU - Swamy, N. Manjunath
AU - Lieberman, Judy
AU - Shankar, Premlata
PY - 2005/8/1
Y1 - 2005/8/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=23044517213&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=23044517213&partnerID=8YFLogxK
U2 - 10.1182/blood-2004-10-3959
DO - 10.1182/blood-2004-10-3959
M3 - Article
C2 - 15831707
AN - SCOPUS:23044517213
VL - 106
SP - 818
EP - 826
JO - Blood
JF - Blood
SN - 0006-4971
IS - 3
ER -