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Woodchuck hepatitis virus post-transcriptional regulatory element deleted from X protein and promoter sequences enhances retroviral vector titer and expression

Gene Therapy volume 13, pages 641–645 (2006)Cite this article

Abstract

Introduction of the post-transcriptional regulatory element (PRE) of woodchuck hepatitis virus (WHV) into the 3′ untranslated region of retroviral and lentiviral gene transfer vectors enhances both titer and transgene expression. Optimal use of the PRE is often necessary to obtain vectors with sufficient performance for therapeutic applications. The enhancing activity of the PRE depends on the precise configuration of its sequence and the context of the vector and cell into which it is introduced. However, data obtained in the context of WHV-associated hepatocellular carcinomas suggests that the PRE might potentially contribute to tumorigenesis, especially if encoding a truncated version of the WHV X protein. Oncogenic side effects of lentiviral vectors containing the PRE have reinforced these safety concerns, although a causal role of the PRE remained unproven. Here, we demonstrate that PRE mutants can be generated that are devoid of X protein open reading frames (ORFs) as well as other ORFs exceeding 25 amino acids, without significant loss of RNA enhancement activity. Furthermore, the X protein promoter could be deleted without compromising the enhancement of vector titers and transgene expression. Such a modified PRE sequence appears useful for future vector design.

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Acknowledgements

We thank R Seyd and M Id for excellent technical assistance. We would like to thank H Schaller, U Protzer and H-G Kräusslich for helpful suggestions. This work was supported by Grants of the European Union (CONSERT, LSHB-CT-2004-005242) and the Deutsche Forschungsgemeinschaft (DFG BA 1837/4).

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  1. A Schambach and J Bohne: These two authors contributed equally to this work.

Authors and Affiliations

  1. Department of Hematology, Hemostaseology and Oncology, Hannover Medical School, Hannover, Germany

    A Schambach, J Bohne & C Baum

  2. Division of Experimental Hematology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA

    C Baum

  3. Applied Virology and Gene Therapy, Georg-Speyer-Haus, Frankfurt, Germany

    F G Hermann, L Egerer, D von Laer & T Giroglou

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  1. A Schambach
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  2. J Bohne
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  7. T Giroglou
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Correspondence to T Giroglou.

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Schambach, A., Bohne, J., Baum, C. et al. Woodchuck hepatitis virus post-transcriptional regulatory element deleted from X protein and promoter sequences enhances retroviral vector titer and expression. Gene Ther 13, 641–645 (2006). https://doi.org/10.1038/sj.gt.3302698

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