In vitro and in vivo characterization of a second functional hairpin ribozyme against HIV-1

Mang Yu, Eric Poeschla, Osamu Yamada, Paula Degrandis, Mark C. Leavitt, Marina Heusch, Jiing Kuala Yees, Flossie Wong-Stahl, Arnold Hampel

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Abstract

We have constructed a hairpin ribozyme targeted to cleave a conserved sequence in the HIV-1 pol gene. The ribozymewas modified to include a structure-stabilizing tetraloop. In vitro studies revealed a cleavage efficiency unprecedented for hairpin ribozymes (kcat/Km = 75 min-1 μM-1. Stable retroviral vector transduction of this ribozyme gene in T-cell lines resulted in long-term ribozyme expression. As compared to control vector transduced T-cells, the pol ribozyme-transduced cells exhibited significant inhibition of different strains of HIV-1 virus production; this protection was greater when ribozyme expression was driven from an internal pol III promoter (adenovirus VA1) than when driven by a pol II promoter (the MMLV LTR). These results further demonstrate the potential of hairpin ribozymes as anti-HIV gene therapy agents and suggest possibilities for employing combinations of independently targeted hairpin ribozymes.

Original languageEnglish (US)
Article number95800530
Pages (from-to)381-386
Number of pages6
JournalVirology
Volume206
Issue number1
DOIs
StatePublished - Jan 1 1995

ASJC Scopus subject areas

  • Virology

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    Yu, M., Poeschla, E., Yamada, O., Degrandis, P., Leavitt, M. C., Heusch, M., Yees, J. K., Wong-Stahl, F., & Hampel, A. (1995). In vitro and in vivo characterization of a second functional hairpin ribozyme against HIV-1. Virology, 206(1), 381-386. [95800530]. https://doi.org/10.1016/S0042-6822(95)80053-0