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Bidirectional promoter interference between two widely used internal heterologous promoters in a late-generation lentiviral construct

Gene Therapy volume 15, pages 384–390 (2008)Cite this article

Abstract

Gene transfer vectors encoding two or more genes are potentially powerful research tools and are poised to play an increasingly important role in gene therapy applications. Common strategies employed to express more than one transgene per vector include the use of multiple promoters, internal ribosome entry site (IRES) elements, splicing signals and fusion proteins. Of these, the IRES elements and multiple promoters have been most widely used. The use of multiple promoters, however, may be compromised by interference between promoters, promoter silencing and vector rearrangements or deletions. In this study, we demonstrate promoter interference between two internal heterologous promoters in the context of a late-generation lentiviral vector. The interference, involving the human cytomegalovirus-immediate-early promoter and human elongation-factor-1α promoter, occurred bidirectionally with both promoters markedly impairing expression of the adjacent transcription unit. The data presented not only highlight the potential for interference between these widely-used promoters, but also the value of a sequential approach to vector construction that allows such effects to be recognized.

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Acknowledgements

We thank Professor Inder Verma (Salk Institute, San Diego, USA) for providing the lentiviral vector reagents used in this study. This work was supported by an ASTRA fellowship from the Royal Australasian Collage of Physicians and by the Haemophilia Foundation of Australia. Julie A Curtin was the recipient of a NHMRC post-graduate research scholarship and a CHW New Investigator Grant. We also thank ACCO Australia, for funding support. Samantha L Ginn is the recipient of a fellowship honouring the memory of Noel Dowling.

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Authors and Affiliations

  1. Gene Therapy Research Unit, The Children's Hospital at Westmead and Children's Medical Research Institute, Westmead, New South Wales, Australia

    J A Curtin, A P Dane, A Swanson, I E Alexander & S L Ginn

  2. Discipline of Paediatrics and Child Health, The University of Sydney, Westmead, New South Wales, Australia

    J A Curtin & I E Alexander

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  1. J A Curtin
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Correspondence to I E Alexander.

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Curtin, J., Dane, A., Swanson, A. et al. Bidirectional promoter interference between two widely used internal heterologous promoters in a late-generation lentiviral construct. Gene Ther 15, 384–390 (2008). https://doi.org/10.1038/sj.gt.3303105

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