Virion encapsidation of tRNA3(Lys)-Ribozyme chimeric RNAs inhibits HIV infection

Shawn Westaway, Laurence Cagnon, Zongli Chang, Shirley Li, Haitang Li, Garry P. Larson, John A. Zaia, John J. Rossi

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Retroviruses require a specific host cellular tRNA primer for initiation of first-strand DNA synthesis. This primer is bound by viral proteins and copackaged into virions. We have exploited this property in the design and testing of an antiviral ribozyme fused to tRNA3(Lys), the primer used for lentiviral replication, including human immunodeficiency virus (HIV-1 and HIV-2). The chimera consists of tRNA3(Lys) covalently attached to a hammerhead ribozyme, which is targeted to the region immediately upstream of the primer binding site of the HIV-1 genome. The tRNA-ribozyme chimeric transcript is catalytically active in vitro and is efficiently bound by HIV reverse transcriptase with an affinity similar to that of tRNA3(Lys). We have expressed the chimeric RNAs from either the tRNA3(Lys) intragenic RNA polymerase III promoter or from a human U6 snRNA promoter. The U6 promoter results in up to 10-fold enhanced expression of the tRNA-ribozyme. Most importantly, the tRNA3(Lys)-ribozymes are encapsidated in HIV-1 virions such that they are effective in substantially reducing the level of infectious virus produced from cells cotransfected with HIV-1 proviral DNA. These results demonstrate the feasibility of using this novel strategy to reduce HIV infectivity and more generally indicate the potential power of using the retroviral primer tRNAs as tools for expressing and delivering ribozymes and other antiretroviral RNAs to the virion capsid.

Original languageEnglish (US)
Pages (from-to)185-197
Number of pages13
JournalAntisense and Nucleic Acid Drug Development
Volume8
Issue number3
StatePublished - 1998
Externally publishedYes

Fingerprint

Catalytic RNA
Virion
HIV Infections
HIV-1
Transfer RNA
RNA
Viruses
RNA Polymerase III
HIV Reverse Transcriptase
HIV-2
Capsid
DNA
Viral Proteins
Retroviridae
Antiviral Agents
Genes
Binding Sites
HIV
Genome
Testing

ASJC Scopus subject areas

  • Genetics
  • Pharmacology

Cite this

Westaway, S., Cagnon, L., Chang, Z., Li, S., Li, H., Larson, G. P., ... Rossi, J. J. (1998). Virion encapsidation of tRNA3(Lys)-Ribozyme chimeric RNAs inhibits HIV infection. Antisense and Nucleic Acid Drug Development, 8(3), 185-197.

Virion encapsidation of tRNA3(Lys)-Ribozyme chimeric RNAs inhibits HIV infection. / Westaway, Shawn; Cagnon, Laurence; Chang, Zongli; Li, Shirley; Li, Haitang; Larson, Garry P.; Zaia, John A.; Rossi, John J.

In: Antisense and Nucleic Acid Drug Development, Vol. 8, No. 3, 1998, p. 185-197.

Research output: Contribution to journalArticle

Westaway, S, Cagnon, L, Chang, Z, Li, S, Li, H, Larson, GP, Zaia, JA & Rossi, JJ 1998, 'Virion encapsidation of tRNA3(Lys)-Ribozyme chimeric RNAs inhibits HIV infection', Antisense and Nucleic Acid Drug Development, vol. 8, no. 3, pp. 185-197.
Westaway, Shawn ; Cagnon, Laurence ; Chang, Zongli ; Li, Shirley ; Li, Haitang ; Larson, Garry P. ; Zaia, John A. ; Rossi, John J. / Virion encapsidation of tRNA3(Lys)-Ribozyme chimeric RNAs inhibits HIV infection. In: Antisense and Nucleic Acid Drug Development. 1998 ; Vol. 8, No. 3. pp. 185-197.
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