Cagnon L, Rossi J; Keystone Symposia on Molecular and Cellular Biology: 1998 HIV Pathogenesis and Treatment.
Keyst Symp Mol Cell Biol Keyst Symp Mol Cell Biol. 1998 Mar 13-19; 47 (abstract no. 2004).
Beckman Research Institute of the City of Hope, Duarte, CA.
We developing a multivalent RNA-based strategy to inhibit HIV replication. We have already demonstrated strong HIV inhibition by ribozymes, Rev binding aptamers, tRNAlys3-ribozyme chimeric RNAs and VA1-antisense. These approaches target viral steps which can possibly mutate. Since the CCR-5 beta-chemokine receptor acts as a co-receptor for M-tropic HIV-1 infection, appears to be essential for HIV life cycle, but is a non-essential human gene, it offers a new and promising target for HIV gene therapy. The alteration of its expression should not affect the physiological function of the modified cells but only interfere with their HIV susceptibility. In order to improve our multivalent RNA based strategy to inhibit HIV infection, we have designed a new hammerhead ribozyme, specifically targeted against the CCR-5 HIV-1 co-receptor. Two RNA polymerase III based cassettes, using the U6snRNA promoter and the Adenoviral VA1 gene, have been developed to express this ribozyme. A RNA Polymerase II transcript is also obtained from the 5'LTR of the pG1Na retroviral vector. Both the U6 and VA1 ribozyme constructs are able to cleave their substrate and are highly expressed in cells. The downregulation of the CCR-5 receptor is under evaluation in several cell lines expressing the various constructs. Stably transfected monocytic cell-lines, expressing the different CCR-5 ribozymes will be challenged with HIV-1 M-tropic strains. Once we have defined the best expression (in terms of stability, level of transcription and cell-compartmentalization) and inhibition systems for the CCR-5 ribozyme, the CCR-5 ribozyme cassette will be used in combination with our different anti viral RNAs to transduce PBL and CD34+ cells.
Publication Types:
Keywords:
- Acquired Immunodeficiency Syndrome
- Adenoviridae
- Cell Line
- Chemokines, CC
- Gene Therapy
- HIV Infections
- HIV Seropositivity
- HIV-1
- Humans
- RNA, Catalytic
- RNA, Viral
- Receptors, CCR5
- hammerhead ribozyme
Other ID:
UI: 102236721
From Meeting Abstracts