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Suppression of oncogenic viral interferon regulatory factor (vIRF) of Kaposi's sarcoma–associated herpesvirus by ribozyme-mediated cleavage

Cancer Gene Therapy volume 8, pages 285–293 (2001)Cite this article

Abstract

Kaposi's sarcoma–associated herpesvirus/human herpesvirus 8 (KSHV/HHV8) has been etiologically associated with several malignancies including Kaposi's sarcoma and primary effusion lymphoma. Oncogenic viral interferon regulatory factor (vIRF) encoded by KSHV ORF-K9 is a homologue of cellular interferon regulatory factor (IRF), and has been demonstrated to inhibit type I/II interferon signal transduction and transform NIH3T3 cells through the interactions with IRF-1, IRF-3, and CBP/p300 proteins. To counteract vIRF's pathogenic role, we have developed five ribozymes targeting ORF-K9 mRNA to suppress vIRF expression. The vIRF RNA substrates were cleaved up to 80% in a substrate-specific manner in transcript cleavage assays in vitro. In a transient transfection assay, two of the ribozymes efficiently suppressed the expression of vIRF protein measured by dual-color immunofluorescence assay that simultaneously detects the expression of both vIRF protein and ribozyme. Flow cytometry analysis showed that these ribozymes reduced vIRF expression up to 76%. A mutant ribozyme had no cleavage activity in vitro, but exhibited antisense effect in vivo. These results suggest that the ribozymes may provide a new approach for functional knockout of vIRF gene, and are potential candidates of antiviral therapy for KSHV-related malignancies. Cancer Gene Therapy (2001) 8, 285–293

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Acknowledgements

We thank Dr. Arun K. Roy for his kind suggestions. This work was supported, in part, by the Howard Hughes Medical Institute through the University of Texas Health Science Center at San Antonio for the Research Resources Program for Medical Schools S/G 7, Elsa U. Pardee Foundation, Association for International Cancer Research, and NIH Grant HL60604-01 to S.-J. Gao.

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Authors and Affiliations

  1. Department of Pediatrics, The University of Texas Health Science Center at San Antonio, San Antonio, 78229, Texas

    Yan-Jin Zhang, Xin-Ping Wang, Jian-Hong Deng & Shou-Jiang Gao

  2. Department of Microbiology, The University of Texas Health Science Center at San Antonio, San Antonio, 78229, Texas

    Yan-Jin Zhang, Xin-Ping Wang, Jian-Hong Deng & Shou-Jiang Gao

  3. Cancer Therapy and Research Center, San Antonio, 78229, Texas

    Richard A Salinas & Noboru Oishi

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  1. Yan-Jin Zhang
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Correspondence to Shou-Jiang Gao.

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Zhang, YJ., Wang, XP., Deng, JH. et al. Suppression of oncogenic viral interferon regulatory factor (vIRF) of Kaposi's sarcoma–associated herpesvirus by ribozyme-mediated cleavage. Cancer Gene Ther 8, 285–293 (2001). https://doi.org/10.1038/sj.cgt.7700299

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