Intravirion targeting of a functional anti-human immunodeficiency virus ribozyme directed to pol

Virology. 2000 Feb 15;267(2):174-84. doi: 10.1006/viro.1999.0112.

Abstract

Ribozymes are catalytic RNAs that offer several advantages as specific therapeutic genes against human immunodeficiency virus type 1 (HIV-1). Significant challenges in antiviral uses of ribozymes include (1) how best to express and to deliver this agent and (2) what is the best locale to target ribozymes against HIV-1 RNA. To explore the former, we have previously characterized several vector systems for efficient expression/delivery of anti-HIV-1 ribozymes (Dropulic et al., 1992; Dropulic and Jeang, 1994a; Smith et al., 1997). Here, to investigate an optimal locale for ribozyme-targeting, we asked whether it might be advantageous to direct ribozymes into HIV-1 virions as opposed to the more conventional approach of targeting ribozymes into infected cells. Two series of experiments were performed. First, we demonstrated that anti-HIV-1 ribozymes could indeed be packaged specifically and efficiently into virions. Second, we compared the virus suppressing activity of a packageable ribozyme with its counterpart, which cannot be packaged into HIV-1 virions. Our results showed that although both ribozymes cleaved HIV-1 genomic RNA in vitro with equivalent efficiencies, the former ribozyme demonstrated significantly higher virus-suppressing activity than the latter. These findings provide proof-of-principle that to combat productive HIV-1 replication, intravirion targeting is more effective than intracellular targeting of ribozymes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Cell Line
  • Gene Products, pol / genetics*
  • Gene Products, pol / metabolism
  • Gene Targeting
  • HIV / genetics*
  • HIV / metabolism
  • HeLa Cells
  • Humans
  • Hydrolysis
  • RNA, Catalytic / genetics*
  • RNA, Catalytic / metabolism
  • RNA, Viral / genetics
  • RNA, Viral / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Virion / genetics*
  • Virion / physiology
  • Virus Assembly / genetics

Substances

  • Gene Products, pol
  • RNA, Catalytic
  • RNA, Viral