Enhanced gene silencing of HIV-1 specific siRNA using microRNA designed hairpins

Daniel Boden; Oliver Pusch; Rebecca Silbermann; Fred Lee; Lynne Tucker; Bharat Ramratnam

doi:10.1093/nar/gkh278

Enhanced gene silencing of HIV-1 specific siRNA using microRNA designed hairpins

Daniel Boden, Oliver Pusch, Rebecca Silbermann, Fred Lee, Lynne Tucker, Bharat Ramratnam

Research output: Contribution to journal › Article › peer-review

212 Scopus citations

Abstract

Post-transcriptional inhibition of HIV-1 replication can be achieved by RNA interference (RNAi). The cellular expression of short interfering RNA (siRNA) or short hairpin RNA (shRNA) homologous to regions of the HIV-1 genome decreases viral replication by the selective degradation of targeted RNA. Here, we demonstrate that another class of noncoding regulatory RNA, termed microRNA (miRNA), can be used to deliver antiviral RNAi. By incorporating sequences encoding siRNA targeting the HIV-1 transactivator protein tat into a human miR-30 pre-microRNA (pre-miRNA) backbone, we were able to express tat siRNA in cells. The tat siRNA delivered as pre-miRNA precursor was 80% more effective in reducing HIV-1 p24 antigen production than tat siRNA expressed as conventional shRNA. Our results confirm the utility of expressing HIV-1 specific siRNA through a miR-30 precursor stem-loop structure and suggest that this strategy can be used to increase the antiviral potency of RNAi.

Original language	English (US)
Pages (from-to)	1154-1158
Number of pages	5
Journal	Nucleic acids research
Volume	32
Issue number	3
DOIs	https://doi.org/10.1093/nar/gkh278
State	Published - 2004
Externally published	Yes

ASJC Scopus subject areas

Genetics

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10.1093/nar/gkh278

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title = "Enhanced gene silencing of HIV-1 specific siRNA using microRNA designed hairpins",

abstract = "Post-transcriptional inhibition of HIV-1 replication can be achieved by RNA interference (RNAi). The cellular expression of short interfering RNA (siRNA) or short hairpin RNA (shRNA) homologous to regions of the HIV-1 genome decreases viral replication by the selective degradation of targeted RNA. Here, we demonstrate that another class of noncoding regulatory RNA, termed microRNA (miRNA), can be used to deliver antiviral RNAi. By incorporating sequences encoding siRNA targeting the HIV-1 transactivator protein tat into a human miR-30 pre-microRNA (pre-miRNA) backbone, we were able to express tat siRNA in cells. The tat siRNA delivered as pre-miRNA precursor was 80% more effective in reducing HIV-1 p24 antigen production than tat siRNA expressed as conventional shRNA. Our results confirm the utility of expressing HIV-1 specific siRNA through a miR-30 precursor stem-loop structure and suggest that this strategy can be used to increase the antiviral potency of RNAi.",

author = "Daniel Boden and Oliver Pusch and Rebecca Silbermann and Fred Lee and Lynne Tucker and Bharat Ramratnam",

note = "Funding Information: We acknowledge the AIDS Research and Reference Reagent Program Division of AIDS, NIAID, NIH for supplying HIVNL4-3 (contributed by Dr Malcolm Martin). This work was supported by a Clinical Scientist Development Award from the Doris Duke Charitable Foundation, a Daland Fellowship in Clinical Investigation from the American Philosophical Society and a Career Development Grant from NIAID/NIH (B.R.). We thank the Lifespan/Tufts/Brown Retrovirology Core Laboratory and the Center for Cancer Research Development (NIHP20RR017695) for assay support.",

year = "2004",

doi = "10.1093/nar/gkh278",

language = "English (US)",

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AU - Boden, Daniel

AU - Pusch, Oliver

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AU - Lee, Fred

AU - Tucker, Lynne

AU - Ramratnam, Bharat

N1 - Funding Information: We acknowledge the AIDS Research and Reference Reagent Program Division of AIDS, NIAID, NIH for supplying HIVNL4-3 (contributed by Dr Malcolm Martin). This work was supported by a Clinical Scientist Development Award from the Doris Duke Charitable Foundation, a Daland Fellowship in Clinical Investigation from the American Philosophical Society and a Career Development Grant from NIAID/NIH (B.R.). We thank the Lifespan/Tufts/Brown Retrovirology Core Laboratory and the Center for Cancer Research Development (NIHP20RR017695) for assay support.

PY - 2004

Y1 - 2004

N2 - Post-transcriptional inhibition of HIV-1 replication can be achieved by RNA interference (RNAi). The cellular expression of short interfering RNA (siRNA) or short hairpin RNA (shRNA) homologous to regions of the HIV-1 genome decreases viral replication by the selective degradation of targeted RNA. Here, we demonstrate that another class of noncoding regulatory RNA, termed microRNA (miRNA), can be used to deliver antiviral RNAi. By incorporating sequences encoding siRNA targeting the HIV-1 transactivator protein tat into a human miR-30 pre-microRNA (pre-miRNA) backbone, we were able to express tat siRNA in cells. The tat siRNA delivered as pre-miRNA precursor was 80% more effective in reducing HIV-1 p24 antigen production than tat siRNA expressed as conventional shRNA. Our results confirm the utility of expressing HIV-1 specific siRNA through a miR-30 precursor stem-loop structure and suggest that this strategy can be used to increase the antiviral potency of RNAi.

AB - Post-transcriptional inhibition of HIV-1 replication can be achieved by RNA interference (RNAi). The cellular expression of short interfering RNA (siRNA) or short hairpin RNA (shRNA) homologous to regions of the HIV-1 genome decreases viral replication by the selective degradation of targeted RNA. Here, we demonstrate that another class of noncoding regulatory RNA, termed microRNA (miRNA), can be used to deliver antiviral RNAi. By incorporating sequences encoding siRNA targeting the HIV-1 transactivator protein tat into a human miR-30 pre-microRNA (pre-miRNA) backbone, we were able to express tat siRNA in cells. The tat siRNA delivered as pre-miRNA precursor was 80% more effective in reducing HIV-1 p24 antigen production than tat siRNA expressed as conventional shRNA. Our results confirm the utility of expressing HIV-1 specific siRNA through a miR-30 precursor stem-loop structure and suggest that this strategy can be used to increase the antiviral potency of RNAi.

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Enhanced gene silencing of HIV-1 specific siRNA using microRNA designed hairpins

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