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PMID: 2062653 Published · ppublish English Journal Article

Inhibition of HIV-LTR gene expression by oligonucleotides targeted to the TAR element.

Nucleic acids research ·Vol. 19 ·No. 12 ·1991-06-25 ·Pages 3359-68

Vickers T, Baker BF, Cook PD, Zounes M, Buckheit RW, Germany J, Ecker DJ

Abstract

All human immunodeficiency virus mRNAs contain a sequence known as TAR (trans-activating responsive sequence). The TAR element forms a stable RNA stem-loop structure which binds the HIV tat (trans-activator) protein and mediates increased viral gene expression. In principle, molecules which bind to the TAR RNA structure would inhibit trans-activation by perturbing the native RNA secondary structure. We have constructed a series of phosphodiester and phosphorothioate antisense oligonucleotides which specifically bind to the HIV TAR element. Specific binding to the TAR element was demonstrated in vitro with enzymatically synthesized TAR RNA. The TAR-directed phosphorothioates inhibited trans-activation in a sequence-dependent fashion in a cell culture model using an HIV LTR/human placental alkaline phosphatase gene fusion and tat protein supplied in trans. The molecules also inhibited HIV replication in both acute and chronically infected viral assays, but without sequence specificity. We have constructed a series of vectors consisting of the MMTV promoter and 5'-untranslated region of four different mRNAs, including the TAR region, to study the effect of TAR on gene expression in heterologous systems. The results suggest that, in the absence of the HIV LTR, the TAR element has a repressive effect on gene expression, which is relieved by tat.

MeSH Terms
Alkaline Phosphatase/genetics Base Sequence Cell Line Gene Products, tat/metabolism HIV/drug effects,physiology HIV Long Terminal Repeat/drug effects,genetics Humans Molecular Sequence Data Nucleic Acid Conformation Oligodeoxyribonucleotides/pharmacology Oligonucleotides, Antisense/pharmacology RNA, Messenger/biosynthesis,drug effects RNA, Viral/biosynthesis,drug effects Transcriptional Activation/drug effects Virus Replication/genetics tat Gene Products, Human Immunodeficiency Virus
Chemicals
Gene Products, tat Oligodeoxyribonucleotides Oligonucleotides, Antisense RNA, Messenger RNA, Viral tat Gene Products, Human Immunodeficiency Virus Alkaline Phosphatase
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Vickers T
ISIS Pharmaceuticals, Carlsbad, CA 92008.
Baker B F
Cook P D
Zounes M
Buckheit R W
Germany J
Ecker D J
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/19.12.3359
Published
1991-06-25
Pages
3359-68
Language
English
Region
England
NLM ID
0411011
PMCID
PMC328335
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/19.12.3359 PMC Full Text View journal details
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