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

doi:10.1016/S0042-6822(95)80053-0

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

Molecular Medicine

Research output: Contribution to journal › Article › peer-review

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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 (k_cat/K_m = 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 language	English (US)
Article number	95800530
Pages (from-to)	381-386
Number of pages	6
Journal	Virology
Volume	206
Issue number	1
DOIs	https://doi.org/10.1016/S0042-6822(95)80053-0
State	Published - 1995

ASJC Scopus subject areas

Virology

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

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.",

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

note = "Funding Information: M.Y. is supported by the NIH AIDS fellow training grant. E.P. is supported by an NIH physician scientist award. A.H. and P.D. were supported by NIH Grant AI29870 and the Biotechnology Research and Development Corporation. This work was also supported in part by an NIH grant (SPITAT) to F.W.S.",

year = "1995",

doi = "10.1016/S0042-6822(95)80053-0",

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T1 - In vitro and in vivo characterization of a second functional hairpin ribozyme against HIV-1

AU - Yu, Mang

AU - Poeschla, Eric

AU - Yamada, Osamu

AU - Degrandis, Paula

AU - Leavitt, Mark C.

AU - Heusch, Marina

AU - Yees, Jiing Kuala

AU - Wong-Stahl, Flossie

AU - Hampel, Arnold

N1 - Funding Information: M.Y. is supported by the NIH AIDS fellow training grant. E.P. is supported by an NIH physician scientist award. A.H. and P.D. were supported by NIH Grant AI29870 and the Biotechnology Research and Development Corporation. This work was also supported in part by an NIH grant (SPITAT) to F.W.S.

PY - 1995

Y1 - 1995

N2 - 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.

AB - 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.

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

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